Structure of corneal layers, collagen fibrils, and proteoglycans of tree shrew cornea.

PubMed

Almubrad, Turki; Akhtar, Saeed

2011-01-01

The stroma is the major part of the cornea, in which collagen fibrils and proteoglycans are distributed uniformly. We describe the ultrastructure of corneal layers, collagen fibrils (CF), and proteoglycans (PGs) in the tree shrew cornea. Tree shrew corneas (5, 6, and 10 week old animals) and normal human corneas (24, 25, and 54 years old) were fixed in 2.5% glutaraldehyde containing cuprolinic blue in a sodium acetate buffer. The tissue was processed for electron microscopy. The 'iTEM Olympus Soft Imaging Solutions GmbH' program was used to measure the corneal layers, collagen fibril diameters and proteoglycan areas. The tree shrew cornea consists of 5 layers: the epithelium, Bowman's layer, stroma, Descemet's membrane, and endothelium. The epithelium was composed of squamous cells, wing cells and basal cells. The Bowman's layer was 5.5±1.0 µm thick and very similar to a normal human Bowman's layer. The stroma was 258±7.00 µm thick and consisted of collagen fibril lamellae. The lamellae were interlaced with one another in the anterior stroma, but ran parallel to one another in the middle and posterior stroma. Collagen fibrils were decorated with proteoglycan filaments with an area size of 390 ±438 nm(2). The collagen fibril had a minimum diameter of 39±4.25 nm. The interfibrillar spacing was 52.91±6.07 nm. Within the collagen fibrils, very small electron-dense particles were present. The structure of the tree shrew cornea is very similar to that of the normal human cornea. As is the case with the human cornea, the tree shrew cornea had a Bowman's layer, lamellar interlacing in the anterior stroma and electron-dense particles within the collagen fibrils. The similarities of the tree shrew cornea with the human cornea suggest that it could be a good structural model to use when studying changes in collagen fibrils and proteoglycans in non-genetic corneal diseases, such as ectasia caused after LASIK (laser-assisted in situ keratomileusis).

Evaluating the Effects of Riboflavin/UV-A and Rose-Bengal/Green Light Cross-Linking of the Rabbit Cornea by Noncontact Optical Coherence Elastography

PubMed Central

Singh, Manmohan; Li, Jiasong; Han, Zhaolong; Vantipalli, Srilatha; Liu, Chih-Hao; Wu, Chen; Raghunathan, Raksha; Aglyamov, Salavat R.; Twa, Michael D.; Larin, Kirill V.

2016-01-01

Purpose The purpose of this study was to use noncontact optical coherence elastography (OCE) to evaluate and compare changes in biomechanical properties that occurred in rabbit cornea in situ after corneal collagen cross-linking by either of two techniques: ultraviolet-A (UV-A)/riboflavin or rose-Bengal/green light. Methods Low-amplitude (≤10 μm) elastic waves were induced in mature rabbit corneas by a focused air pulse. Elastic wave propagation was imaged by a phase-stabilized swept source OCE (PhS-SSOCE) system. Corneas were then cross-linked by either of two methods: UV-A/riboflavin (UV-CXL) or rose-Bengal/green light (RGX). Phase velocities of the elastic waves were fitted to a previously developed modified Rayleigh-Lamb frequency equation to obtain the viscoelasticity of the corneas before and after the cross-linking treatments. Micro-scale depth-resolved phase velocity distribution revealed the depth-wise heterogeneity of both cross-linking techniques. Results Under standard treatment settings, UV-CXL significantly increased the stiffness of the corneas by ∼47% (P < 0.05), but RGX did not produce statistically significant increases. The shear viscosities were unaffected by either cross-linking technique. The depth-wise phase velocities showed that UV-CXL affected the anterior ∼34% of the corneas, whereas RGX affected only the anterior ∼16% of the corneas. Conclusions UV-CXL significantly strengthens the cornea, whereas RGX does not, and the effects of cross-linking by UV-CXL reach deeper into the cornea than cross-linking effects of RGX under similar conditions. PMID:27409461

UV cross-linking of donor corneas confers resistance to keratolysis.

PubMed

Arafat, Samer N; Robert, Marie-Claude; Shukla, Anita N; Dohlman, Claes H; Chodosh, James; Ciolino, Joseph B

2014-09-01

The aim of this study was to develop a modified ex vivo corneal cross-linking method that increases stromal resistance to enzymatic degradation for use as a carrier for the Boston keratoprosthesis. Ex vivo cross-linking of human corneas was performed using Barron artificial anterior chambers. The corneas were deepithelialized, pretreated with riboflavin solution (0.1% riboflavin/20% dextran), and irradiated with ultraviolet A (UV-A) light (λ = 370 nm, irradiance = 3 mW/cm) for various durations. The combined effect of UV-A and gamma (γ) irradiation was also assessed using the commercially available γ-irradiated corneal donors. The corneas were then trephined and incubated at 37°C with 0.3% collagenase A solution. The time to dissolution of each cornea was compared across treatments. Deepithelialized corneas (no UV light, no riboflavin) dissolved in 5.8 ± 0.6 hours. Cross-linked corneas demonstrated increased resistance to dissolution, with a time to dissolution of 17.8 ± 2.6 hours (P < 0.0001). The corneal tissues' resistance to collagenase increased with longer UV-A exposure, reaching a plateau at 30 minutes. Cross-linking both the anterior and posterior corneas did not provide added resistance when compared with cross-linking the anterior corneas only (P > 0.05). γ-irradiated corneas dissolved as readily as deepithelialized controls regardless of whether they were further cross-linked (5.6 ± 1.2 hours) or not (6.1 ± 0.6 hours) (P = 0.43). Collagen cross-linking of the deepithelialized anterior corneal surface for 30 minutes conferred optimal resistance to in vitro keratolysis by collagenase A.

Screening for previous refractive surgery in eye bank corneas by using optical coherence tomography.

PubMed

Lin, Roger C; Li, Yan; Tang, Maolong; McLain, Marcy; Rollins, Andrew M; Izatt, Joseph A; Huang, David

2007-06-01

To use optical coherence tomography (OCT) to detect previous refractive surgery in donor corneas. We constructed a tabletop OCT scanner operating at 1310-nm wavelength. Donor corneas at the Cleveland Eye Bank were scanned while sealed within the sterile container immersed in Optisol GS. OCT scanning was performed with 7.6-mm-long lines (512 axial scans) along 8 meridians. Anterior and posterior corneal surfaces were automatically mapped using image processing software that we developed. Curvature was computed from the best parabolic fit in the central 5-mm diameter. Layered analysis of the stromal reflectivity was also performed. Twenty-nine corneas from 19 donors were examined. Five had a history of laser in situ keratomileusis (LASIK). The flap interfaces could not be visualized on slit-lamp or OCT images but were confirmed by histology. The death-to-scan time was 22.1 +/- 11.4 (SD) hours for normal corneas and 100.6 +/- 57.5 hours for LASIK corneas. The anterior surface power was 67.5 +/- 2.5 D in control corneas and 64.5 +/- 2.4 D in LASIK corneas (P = 0.023). There was no significance between the 2 groups in terms of posterior curvature and thickness parameters. The anterior/posterior reflectivity ratio in the central 4-mm diameter was significantly lower in post-LASIK corneas than in control (P < 0.05). OCT provides thickness, topography, and reflectivity maps of donor corneas without taking them out of preservation medium and container. The anterior curvature and the anterior/posterior stromal reflectivity ratio may be useful for detecting previous LASIK.

Imaging of Keratoconic and normal human cornea with a Brillouin imaging system (Conference Presentation)

NASA Astrophysics Data System (ADS)

Besner, Sebastien; Shao, Peng; Scarcelli, Giuliano; Pineda, Roberto; Yun, Seok-Hyun (Andy)

2016-03-01

Keratoconus is a degenerative disorder of the eye characterized by human cornea thinning and morphological change to a more conical shape. Current diagnosis of this disease relies on topographic imaging of the cornea. Early and differential diagnosis is difficult. In keratoconus, mechanical properties are found to be compromised. A clinically available invasive technique capable of measuring the mechanical properties of the cornea is of significant importance for understanding the mechanism of keratoconus development and improve detection and intervention in keratoconus. The capability of Brillouin imaging to detect local longitudinal modulus in human cornea has been demonstrated previously. We report our non-contact, non-invasive, clinically viable Brillouin imaging system engineered to evaluate mechanical properties human cornea in vivo. The system takes advantage of a highly dispersive 2-stage virtually imaged phased array (VIPA) to detect weak Brillouin scattering signal from biological samples. With a 1.5-mW light beam from a 780-nm single-wavelength laser source, the system is able to detect Brillouin frequency shift of a single point in human cornea less than 0.3 second, at a 5μm/30μm lateral/axial resolution. Sensitivity of the system was quantified to be ~ 10 MHz. A-scans at different sample locations on a human cornea with a motorized human interface. We imaged both normal and keratoconic human corneas with this system. Whereas no significantly difference were observed outside keratocnic cones compared with normal cornea, a highly statistically significantly decrease was found in the cone regions.

[Biomechanical condition of the cornea as a new indicator for pathological and structural changes].

PubMed

Spörl, E; Terai, N; Haustein, M; Böhm, A G; Raiskup-Wolf, F; Pillunat, L E

2009-06-01

Several methods permit the measurement of geometric parameters of the cornea, but until now biomechanical conditions of the cornea have been ignored (e.g. in refractive corneal surgery). Besides the geometric condition, biomechanical properties of the cornea have been shown to influence applanation measurement of intra-ocular pressure (IOP) and epidemiological studies have identified corneal thickness as an independent risk factor for the development and progression of glaucoma. The aim of this investigation was to characterize the biomechanical properties of the cornea using the ocular response analyzer (ORA). The ocular response analyzer (ORA) is a new method available for non-contact measurement of the biomechanical properties of the cornea. We evaluated the reproducibility of measurements, the difference between static and dynamic factors and the impact of independent factors (e.g. IOP, age, CCT, swelling of the cornea) on 2,500 measurements of corneal hysteresis (CH) and corneal resistance factor (CRF). In a large sample size we observed changes in CH and CRF after refractive surgery procedures (LASIK, UV-A cross-linking, keratoplasty) and in other corneal disorders (keratoconus, corneal dystrophies). CRF and CH changes may reflect structural changes of the cornea. Thus, the ORA provides valuable information for a better understanding and characterization of the biomechanical condition of the cornea, especially with regard to diseases such as keratoconus and glaucoma.

[Comparative analysis of the donor cornea quality and of the interval between death and preservation before and after new sanitary and technique rules in a University Eye Bank].

PubMed

Zantut, Fabio; Holzchuh, Ricardo; Boni, Reginaldo Carlos; Mackus, Eva Cristina; Zantut, Paulo Roberto; Nakano, Claudio; Netto, Adamo Lui; Hida, Richard Yudi

2012-01-01

To compare the interval between death and enucleation (ΔT-O-E), between enucleation and preservation (ΔT-E-P) and the quality of the cornea before and after the implantation of new technique and sanitary rules. A retrospective study that evaluated the records of cornea donors in Sao Paulo's Santa Casa Eye Tissue Bank 2 years before and 2 years after the implementation new sanitary rules. An increase was observed in the absolute number of 205 to 374 donors following the adopted changes. There was no statistically significant difference in Δt-O-E and ΔT-E-P before and after the implemented changes. Of the total of 1,105 donor corneas, 388 donor corneas were observed before the changes and 717 donor corneas after the implemented changes. We observed a statistically significant increase in grading of donor cornea quality from 1.76 ± 0.90 to 1.94 ± 0.88 after the implementation of new standards of resolution. After the changes required by Resolution 347, there was a large increase in the number of donated, taken and preserved corneas. The BTO has not diminished the ΔT O-E and ΔT E-P. Cornea quality presented itself lower after the new rules.

The influence of intraocular pressure and air jet pressure on corneal contactless tonometry tests.

PubMed

Simonini, Irene; Pandolfi, Anna

2016-05-01

The air puff is a dynamic contactless tonometer test used in ophthalmology clinical practice to assess the biomechanical properties of the human cornea and the intraocular pressure due to the filling fluids of the eye. The test is controversial, since the dynamic response of the cornea is governed by the interaction of several factors which cannot be discerned within a single measurement. In this study we describe a numerical model of the air puff tests, and perform a parametric analysis on the major action parameters (jet pressure and intraocular pressure) to assess their relevance on the mechanical response of a patient-specific cornea. The particular cornea considered here has been treated with laser reprofiling to correct myopia, and the parametric study has been conducted on both the preoperative and postoperative geometries. The material properties of the cornea have been obtained by means of an identification procedure that compares the static biomechanical response of preoperative and postoperative corneas under the physiological IOP. The parametric study on the intraocular pressure suggests that the displacement of the cornea׳s apex can be a reliable indicator for tonometry, and the one on the air jet pressure predicts the outcomes of two or more distinct measurements on the same cornea, which can be used in inverse procedures to estimate the material properties of the tissue. Copyright © 2015 Elsevier Ltd. All rights reserved.

Characterization of Rabbit Corneas Subjected to Stromal Stiffening by the Açaí Extract (Euterpe oleracea).

PubMed

Bersanetti, Patrícia A; Bueno, Tatiane L N; Morandim-Giannetti, Andreia de A; Nogueira, Regina F; Matos, Jivaldo R; Schor, Paulo

2017-04-01

In this study, we characterized rabbit corneas subjected to corneal cross-linking (CXL) with açaí extract compared with a riboflavin photo-stimulated procedure. The corneas of the slaughterhouse rabbits were divided into three groups: control, consisting of untreated corneal samples; riboflavin/UVA, where corneas were treated with 0.1% riboflavin photo-stimulated at 365 nm as the standard protocol; and açaí, where the samples were subjected to 4% açaí extract for 0.5-2 h. After the CXL procedure, corneas of the three groups were characterized by analyzing their elastic modulus and thermal denaturation profile. The elastic modulus at 3% strain showed an approximately threefold increase in the riboflavin/UVA group and 10.5 times in the corneas treated with 4% açaí extract for 2 h, compared with the control group (p < 0.01). The denaturation temperature values of the two groups of crosslinked corneas increased significantly (p < 0.05) and were more pronounced in the açaí group. The açaí extract was effective in promoting CXL in rabbit corneas as characterized by the different techniques.

In vivo confocal microscopy of human cornea covered with human amniotic membrane.

PubMed

Mimura, Tatsuya; Yamagami, Satoru; Usui, Tomohiko; Honda, Norihiko; Araki, Fumiyuki; Amano, Shiro

2008-01-01

Amniotic membrane transplantation has been widely performed to reconstruct the surface of the eye and treat chemical burns or epithelial defects. However, we have difficulty observing the cornea through the opaque transplanted amniotic membrane by slit-lamp biomicroscopy. We investigated the use of confocal microscopy for observation of human corneas covered with amniotic membrane. Human amniotic membrane was placed onto the normal corneas of five volunteers aged 22-24 years. Then, all layers of the covered corneas were observed by in vivo confocal microscopy. Confocal microscopy displayed the epithelium, basement membrane, and stroma of the amniotic membrane. It also displayed the corneal epithelium. Furthermore, corneal stromal keratocytes and the corneal endothelium were clearly observed through the amniotic membrane by confocal microscopy. We demonstrated that in vivo confocal microscopy enabled us to observe all layers of corneas covered with amniotic membrane in normal human eyes. Our findings suggest that confocal microscopy may have advantages for clinical examination of the ocular surface, including all layers of the cornea.

SPECIAL ISSUE DEVOTED TO MULTIPLE RADIATION SCATTERING IN RANDOM MEDIA: Optical coherent tomography measurements of the diffusion rate of water and drugs in an isolated and whole cornea

NASA Astrophysics Data System (ADS)

Larin, Kirill V.; Ghosn, M. G.

2006-12-01

The passive diffusion of drugs through the epithelial surfaces of an eye (the most widespread method for medical treatment of various diseases) is considered. The permeability of water and drugs through rabbit cornea was measured in the isolated cornea (separate from an eye) and in the whole cornea. The permeability coefficients of water and dexamethasone were estimated by the method of optical coherence tomography (OCT). Because multiple photon scattering introduces noise and distortions to the OCT signal, measurements were performed at depths up to 500 μm where most likely single scattering of light occurs in cornea. It is shown that the permeability coefficients in the isolated and whole cornea strongly differ from each other. For example, the water permeability in the isolated and whole cornea is (7.09±0.12)×10-5 and (1.71±0.51)×10-5 cm s-1, respectively.

Clinical and epidemiological aspects of cornea transplant patients of a reference hospital 1

PubMed Central

Cruz, Giovanna Karinny Pereira; de Azevedo, Isabelle Campos; Carvalho, Diana Paula de Souza Rego Pinto; Vitor, Allyne Fortes; Santos, Viviane Euzébia Pereira; Ferreira, Marcos Antonio

2017-01-01

ABSTRACT Objective: clinically characterizing cornea transplant patients and their distribution according to indicated and post-operative conditions of cornea transplantation, as well as estimating the average waiting time. Method: a cross-sectional, descriptive and analytical study performed for all cornea transplants performed at a reference service (n=258). Data were analyzed using Statistical Package for the Social Sciences, version 20.0. Results: the main indicator for cornea transplant was keratoconus. The mean waiting time for the transplant was approximately 5 months and 3 weeks for elective transplants and 9 days for urgent cases. An association between the type of corneal disorder with gender, age, previous surgery, eye classification, glaucoma and anterior graft failure were found. Conclusion: keratoconus was the main indicator for cornea transplant. Factors such as age, previous corneal graft failure (retransplantation), glaucoma, cases of surgeries prior to cornea transplant (especially cataract surgery) may be related to the onset corneal endothelium disorders. PMID:28614429

[The 2009 performance report of the German cornea banks].

PubMed

Schrage, N; Reinhard, T; Seitz, B; Hermel, M; Böhringer, D; Reinshagen, H

2011-03-01

In Germany, human tissue for corneal and amniotic transplantation is supplied by 27 cornea banks. The Section for Tissue Transplantation and Biotechnology of the German Ophthalmological Society records the cornea banks' activities by means of an annual questionnaire. In 2009, a total of 4,818 corneal grafts were processed by 21 responding cornea banks, and 57% were deemed suitable for transplantation. This ratio is slightly higher than the European average. In addition, German cornea banks released 1,257 amniotic grafts in 2009. German cornea banks are currently facing new regulatory issues due to updated legislation regarding tissue transplantation. Recent updates in European law have limited the cutoff time for postmortem blood sampling to 24 h, and this regulation may lead to a significant reduction in potential donors.

Atomic force microscopy analysis of human cornea surface after UV (λ=266 nm) laser irradiation

NASA Astrophysics Data System (ADS)

Spyratou, E.; Makropoulou, M.; Moutsouris, K.; Bacharis, C.; Serafetinides, A. A.

2009-07-01

Efficient cornea reshaping by laser irradiation for correcting refractive errors is still a major issue of interest and study. Although the excimer laser wavelength of 193 nm is generally recognized as successful in ablating corneal tissue for myopia correction, complications in excimer refractive surgery leads to alternative laser sources and methods for efficient cornea treatment. In this work, ablation experiments of human donor cornea flaps were conducted with the 4th harmonic of an Nd:YAG laser, with different laser pulses. AFM analysis was performed for examination of the ablated cornea flap morphology and surface roughness.

Structure of corneal layers, collagen fibrils, and proteoglycans of tree shrew cornea

PubMed Central

Almubrad, Turki

2011-01-01

Purpose The stroma is the major part of the cornea, in which collagen fibrils and proteoglycans are distributed uniformly. We describe the ultrastructure of corneal layers, collagen fibrils (CF), and proteoglycans (PGs) in the tree shrew cornea. Methods Tree shrew corneas (5, 6, and 10 week old animals) and normal human corneas (24, 25, and 54 years old) were fixed in 2.5% glutaraldehyde containing cuprolinic blue in a sodium acetate buffer. The tissue was processed for electron microscopy. The ‘iTEM Olympus Soft Imaging Solutions GmbH’ program was used to measure the corneal layers, collagen fibril diameters and proteoglycan areas. Results The tree shrew cornea consists of 5 layers: the epithelium, Bowman’s layer, stroma, Descemet’s membrane, and endothelium. The epithelium was composed of squamous cells, wing cells and basal cells. The Bowman’s layer was 5.5±1.0 µm thick and very similar to a normal human Bowman’s layer. The stroma was 258±7.00 µm thick and consisted of collagen fibril lamellae. The lamellae were interlaced with one another in the anterior stroma, but ran parallel to one another in the middle and posterior stroma. Collagen fibrils were decorated with proteoglycan filaments with an area size of 390 ±438 nm2. The collagen fibril had a minimum diameter of 39±4.25 nm. The interfibrillar spacing was 52.91±6.07 nm. Within the collagen fibrils, very small electron-dense particles were present. Conclusions The structure of the tree shrew cornea is very similar to that of the normal human cornea. As is the case with the human cornea, the tree shrew cornea had a Bowman's layer, lamellar interlacing in the anterior stroma and electron-dense particles within the collagen fibrils. The similarities of the tree shrew cornea with the human cornea suggest that it could be a good structural model to use when studying changes in collagen fibrils and proteoglycans in non-genetic corneal diseases, such as ectasia caused after LASIK (laser-assisted in situ keratomileusis). PMID:21921979

Cost Minimization Analysis of Precut Cornea Grafts in Descemet Stripping Automated Endothelial Keratoplasty

PubMed Central

Yong, Kai-Ling; Nguyen, Hai V.; Cajucom-Uy, Howard Y.; Foo, Valencia; Tan, Donald; Finkelstein, Eric A.; Mehta, Jodhbir S.

2016-01-01

Abstract Descemet stripping automated endothelial keratoplasty (DSAEK) is the most common corneal transplant procedure. A key step in the procedure is preparing the donor cornea for transplantation. This can be accomplished via 1 of 3 alternatives: surgeon cuts the cornea on the day of surgery, the cornea is precut ahead of time in an offsite facility by a trained technician, or a precut cornea is purchased from an eye bank. Currently, there is little evidence on the costs and effectiveness of these 3 strategies to allow healthcare providers decide upon the preferred method to prepare grafts. The aim of this study was to compare the costs and relative effectiveness of each strategy. The Singapore National Eye Centre and Singapore Eye Bank performed both precut cornea and surgeon-cut cornea transplant services between 2009 and 2013. This study included 110 subjects who received precut cornea and 140 who received surgeon-cut cornea. Clinical outcomes and surgical duration were compared across the strategies using the propensity score matching. The cost of each strategy was estimated using the microcosting and consisted of facility costs and procedural costs including surgical duration. One-way sensitivity analysis and threshold analysis were performed. The cost for DSAEK was highest for the surgeon-cut approach ($13,965 per procedure), followed by purchasing precut corneas ($12,659) and then setting up precutting ($12,421). The higher procedural cost of the surgeon-cut approach was largely due to the longer duration of the procedure (surgeon-cut = 72.54 minutes, precut = 59.45 minutes, P < 0.001) and the higher surgeon fees. There was no evidence of differences in clinical outcomes between grafts that were precut or surgeon-cut. Threshold analysis demonstrated that if the number of cases was below 31 a year, the strategy that yielded the lowest cost was purchasing precut cornea from eye bank. If there were more than 290 cases annually, the cheapest option would be to setup precutting facility. Our findings suggest that it is more efficient for centers that are performing a large number of cornea transplants (more than 290 cases) to set up their own facility to conduct precutting. PMID:26937927

Zinc oxide tetrapods inhibit herpes simplex virus infection of cultured corneas

PubMed Central

Duggal, Neil; Jaishankar, Dinesh; Yadavalli, Tejabhiram; Hadigal, Satvik; Mishra, Yogendra Kumar; Adelung, Rainer

2017-01-01

Purpose Infection of the human cornea by herpes simplex virus type-1 (HSV-1) can cause significant vision loss. The purpose of this study was to develop an ex vivo model to visualize viral growth and spread in the cornea. The model was also used to analyze cytokine production and study the antiviral effects of zinc oxide tetrapods. Methods A β-galactosidase-expressing recombinant virus, HSV-1(KOS)tk12, was used to demonstrate the ability of the virus to enter and develop blue plaques on human corneal epithelial (HCE) cells and corneal tissues. Freshly obtained porcine corneas were cultured and then scratched before infection with HSV-1(KOS)tk12. The blue plaques on the corneas were imaged using a stereomicroscope. Western blot analysis for HSV-1 proteins was performed to verify HSV-1 infection of the cornea. Using the ex vivo model, zinc oxide tetrapods were tested for their anti-HSV-1 potential, and a cytokine profile was developed to assess the effects of the treatment. Results Cultured corneas and the use of β-galactosidase-expressing HSV-1(KOS)tk12 virus can provide an attractive ex vivo model to visualize and study HSV-1 entry and spread of the infection in tissues. We found that unlike cultured HCE cells, which demonstrated nearly 100% infectivity, HSV-1 infection of the cultured cornea was more restrictive and took longer to develop. We also found that the zinc oxide tetrapod–shaped nano- and microstructures inhibited HSV infection of the cultured cells, as well as the cultured corneas. The cytokine profile of the infected samples was consistent with previous studies of HSV-1 corneal infection. Conclusions The ability to visualize HSV-1 growth and spread in corneal tissues can provide new details about HSV-1 infection of the cornea and the efficacy of new cornea-specific antiviral drug candidates. The ex vivo model also demonstrates antiviral effects of zinc oxide tetrapods and adequately portrays the drug delivery issues that cornea-specific treatments face. PMID:28275313

Comparison of the characteristics in hen and quail corneas as experimental models of refractive surgery.

PubMed

Gonçalves, G C; Pérez-Merino, P; Martínez-García, M C; Barcía, A; Merayo-Loves, J

2016-07-01

To compare the histological, morphological and the biophysical measurements between hen and quail corneas, in order to determine which of them were better suited for use as an animal model for research into corneal refractive surgery. A study was performed using the biophysical measurements of the cornea (curvature, thickness, refraction, and axial length) of 20 animals (10 hens and 10 quails). The corneas were then prepared for histological analysis under microscopy light. The analysis showed that both groups have the same number of corneal layers as the human cornea and with an evident Bowman's layer. The thickness of the hen cornea and axial length of the eye, 225.3±18.4μm and 12.8±0.25mm, respectively, were larger than that of the quail (P<.01 and P<.001, respectively). The radius of curvature for the hen central cornea, 3.65±0.08mm, was greater than that for the quail (P<.001), but the refractive power of each cornea was similar. The proportion of total corneal thickness of the hen stroma, 82.6%, was more similar to that of the human than was the quail stroma, 72.5%. Within the hen stroma, the density of keratocytes, 8.57±1.49 per 5,000μm(2), was about half that in the quail stroma (P<.005). Because of the large size of the hen cornea, the stromal thickness and proportional similarity of the corneal layers with human cornea, the hen maybe better than the quail as an alternative species suitable for use in studies of corneal refractive surgery. Copyright © 2016 Sociedad Española de Oftalmología. Published by Elsevier España, S.L.U. All rights reserved.

Melatonin Entrains PER2::LUC Bioluminescence Circadian Rhythm in the Mouse Cornea

PubMed Central

Baba, Kenkichi; Davidson, Alec J.; Tosini, Gianluca

2015-01-01

Purpose Previous studies have reported the presence of a circadian rhythm in PERIOD2::LUCIFERASE (PER2::LUC) bioluminescence in mouse photoreceptors, retina, RPE, and cornea. Melatonin (MLT) modulates many physiological functions in the eye and it is believed to be one of the key circadian signals within the eye. The aim of the present study was to investigate the regulation of the PER2::LUC circadian rhythm in mouse cornea and to determine the role played by MLT. Methods Corneas were obtained from PER2::LUC mice and cultured to measure bioluminescence rhythmicity in isolated tissue using a Lumicycle or CCD camera. To determine the time-dependent resetting of the corneal circadian clocks in response to MLT or IIK7 (a melatonin type 2 receptor, MT2, agonist) was added to the cultured corneas at different times of the day. We also defined the location of the MT2 receptor within different corneal layers using immunohistochemistry. Results A long-lasting bioluminescence rhythm was recorded from cultured PER2::LUC cornea and PER2::LUC signal was localized to the corneal epithelium and endothelium. MLT administration in the early night delayed the cornea rhythm, whereas administration of MLT at late night to early morning advanced the cornea rhythm. Treatment with IIK7 mimicked the MLT phase-shifting effect. Consistent with these results, MT2 immunoreactivity was localized to the corneal epithelium and endothelium. Conclusions Our work demonstrates that MLT entrains the PER2::LUC bioluminescence rhythm in the cornea. Our data indicate that the cornea may represent a model to study the molecular mechanisms by which MLT affects the circadian clock. PMID:26207312

Effects of Microgravity on Quail Eye Development

NASA Technical Reports Server (NTRS)

Conrad, Gary W.

1996-01-01

During embryonic development, the most exposed tissue of the eye, the cornea, becomes differentially bulged outward because of constant intraocular pressure (IOP). The component cells of the cornea secrete a unique, paracrystalline extracellular matrix (the stroma) composed of orthogonal plies of collagen fibrils and proteoglycans. The cornea remains avascular, becomes transparent, and becomes more densely innervated than any other region on the surface of the body. Corneas from chicken embryos that flew on STS-47 contain many more cellular processes in the outermost region of the stroma (Bowman's Layer) than any corresponding region of control corneas. These processes appear to be cross-sections of cytoplasmic extensions of cells and are found in that region of Bowman's Layer immediately beneath the basal lamina of the corneal epithelium. Here, we propose to compare corneas of quail that flew in space on Mir-1 with those of ground controls to determine if the same unusual cellular processes are seen as in the space-flown chicken corneas. In the central regions of such space-flown corneas, the processes appear to be either portions of basal epithelial cells whose pseudopodial extensions have migrated down through their own basal lamina into the stroma, or corneal nerves that have innervated the corneal stroma in an unusual manner. Eyeballs of embryos fixed on Mir-1, control embryos fixed at KSC and clinostated embryos fixed at KSU, will provide corneas for this study. Electron microscopy will be used to assess the distribution of the cellular processes in Bowman's Layer in the central region of each cornea. Attempts also will be made to determine the relative glycosaminoglycan distributions in the corneal stromas by indirect immunofluorscence and to record whole-mount staining patterns of the corneal nerves.

Finger's amniotic membrane buffer technique: protecting the cornea during radiation plaque therapy.

PubMed

Finger, Paul T

2008-04-01

To use amniotic membranes as a buffer between the cornea and radioactive eye plaques. Six melanomas were treated with ophthalmic plaque radiation therapy. Plaque-tumor localization required that a portion of the gold plaque touch the cornea during treatment. To enhance patient comfort and protect the cornea, an (0.1-mm-thick) amniotic membrane was interposed between the metal plaque edge and the cornea. Minimal ocular discomfort was noted during plaque radiation therapy. On a scale of 1 (none) to 10 (severe), all 6 patients reported pain levels of 1. As a tissue equivalent and because the mean thickness was only 0.1 mm, amniotic membranes had no significant effect on radiation dose calculations. No adverse effects, infections, or abrasions were noted. The amniotic membrane buffer technique improves patient comfort and protects the cornea during ophthalmic plaque radiation therapy.

Identification of biomechanical properties of the cornea: the ocular response analyzer.

PubMed

Terai, Naim; Raiskup, Frederik; Haustein, Michael; Pillunat, Lutz E; Spoerl, Eberhard

2012-07-01

Several methods have been devised for measuring geometric parameters of the cornea but, until now, the biomechanics of the cornea have been largely ignored. The relatively new Ocular Response Analyzer (ORA) provides such biomechanical information. In order to correctly interpret the underlying biomechanics of ORA data, we review reported ORA measurements and provide a compendium of factors influencing these measurements, with discussion of possible explanations for ORA measurement results. This review comprised a literature search using "ocular response analyzer" and "ocular response analyser" as keywords. We reviewed and compared reported results from recent ORA studies so obtained, with an eye to understanding corneal biomechanics. Several ORA biomechanical parameters of the cornea - corneal hysteresis (CH) and corneal resistant factor (CRF) - characterize the viscoelastic properties of the cornea, especially those of the ground substance. The impact on CH and CRF values of various independent factors, e.g. intraocular pressure (IOP), age, central corneal thickness (CCT), and corneal swelling, are discussed. The impact on CH and CRF of treatment-related structural changes of the cornea, i.e. those occurring after refractive surgical procedures, placement of intracorneal rings, and collagen crosslinking (CXL), as well as pathological changes of the cornea, e.g. those resulting from keratoconus, edema, and glaucoma, are discussed. Changes in CRF and CH may be reflective of structural changes in the ground substance of the cornea. Thus, ORA provides invaluable information for delineating biomechanical conditions pertaining to the cornea, with special regard to ocular diseases, e.g. keratoconus and glaucoma.

Comparison of confocal microscopy and two-photon microscopy in mouse cornea in vivo.

PubMed

Lee, Jun Ho; Lee, Seunghun; Gho, Yong Song; Song, In Seok; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

2015-03-01

High-resolution imaging of the cornea is important for studying corneal diseases at cellular levels. Confocal microscopy (CM) has been widely used in the clinic, and two-photon microscopy (TPM) has recently been introduced in various pre-clinical studies. We compared the performance of CM and TPM in normal mouse corneas and neovascularized mouse corneas induced by suturing. Balb/C mice and C57BL/6 mice expressing green fluorescent protein (GFP) were used to compare modalities based on intrinsic contrast and extrinsic fluorescence contrast. CM based on reflection (CMR), CM based on fluorescence (CMF), and TPM based on intrinsic/extrinsic fluorescence and second harmonic generation (SHG) were compared by imaging the same sections of mouse corneas sequentially in vivo. In normal mouse corneas, CMR visualized corneal cell morphologies with some background noise, and CMF visualized GFP expressing corneal cells clearly. TPM visualized corneal cells and collagen in the stroma based on fluorescence and SHG, respectively. However, in neovascularized mouse corneas, CMR could not resolve cells deep inside the cornea due to high background noise from the effects of increased structural irregularity induced by suturing. CMF and TPM visualized cells and induced vasculature better than CMR because both collect signals from fluorescent cells only. Both CMF and TPM had signal decays with depth due to the structural irregularity, with CMF having faster signal decay than TPM. CMR, CMF, and TPM showed different degrees of image degradation in neovascularized mouse corneas. Copyright © 2015 Elsevier Ltd. All rights reserved.

Femtosecond laser cutting of multiple thin corneal stromal lamellae for endothelial bioengineering.

PubMed

Bernard, Aurélien; He, Zhiguo; Forest, Fabien; Gauthier, Anne-Sophie; Peocʼh, Michel; Dumollard, Jean-Marc; Acquart, Sophie; Montard, Romain; Delbosc, Bernard; Gain, Philippe; Thuret, Gilles

2015-02-01

To assess the feasibility of cutting multiple thin stromal lamellae in human donor corneas using a commercial femtosecond laser (FSL) to provide cell carriers for future endothelial graft bioengineering. Eight edematous organ-cultured corneas not suitable for grafting for endothelial reasons were mounted on a Ziemer anterior chamber and cut with a Z6 FSL with 6 successive parallel cuts, from depth to surface. Target thickness of each lamella ranged from 100 to 150 μm depending on initial corneal thickness. Thickness was measured using anterior segment optical coherence tomography before and after cutting on mounted corneas, and on each stromal lamella after detachment. Scanning electron microscopy observation was performed on 4 lamellae and histological cross sections on 1 cornea before detachment. A median of 5 (minimum 3, maximum 7) lamellae was obtained per cornea. All lamellae still attached were the most posterior ones, suggesting that FSL was less efficient because of light scattering by edematous stroma. Cut precision and postdetachment swelling were correlated with anterior-posterior position within the cornea. Median lamella thickness was 127 μm (56-222 μm) before detachment and 196 μm (80-304 μm) after detachment. Surface state was consistent with previously reported FSL lamellar cuts during Descemet stripping automated endothelial keratoplasty. Up to 7 thin lamellae can be cut in stored corneas with an FSL. This method, once optimized primarily by using deswelled, more transparent corneas, could prove effective for recycling unsuitable donor corneas in corneal bioengineering processes.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ediger, M.N.

The laser-induced fluorescence spectrum of rabbit cornea irradiated at ablative intensities was measured. This system directly measured the radiant exposure of fluorescence transmitted through the cornea when the anterior surface of the cornea was irradiated by an ArF excimer laser. Evidence of changing spectral characteristics as a function of total laser dose suggests photochemical changes in the cornea may be occurring. Results are compared with previous data of laser-induced fluorescence in other models and detection schemes.

Collagen organization in the chicken cornea and structural alterations in the retinopathy, globe enlarged (rge) phenotype--an X-ray diffraction study.

PubMed

Boote, Craig; Hayes, Sally; Jones, Simon; Quantock, Andrew J; Hocking, Paul M; Inglehearn, Chris F; Ali, Manir; Meek, Keith M

2008-01-01

An investigation into the collagenous structure of the mature avian cornea is presented. Wide-angle X-ray diffraction is employed to assess collagen organization in 9-month-old chicken corneas. The central 2-4mm corneal region features a preponderance of fibrils directed along the superior-inferior and nasal-temporal orthogonal meridians. More peripherally the orientation of fibrils alters in favor of a predominantly tangential arrangement. The chicken cornea appears to be circumscribed by an annulus of fibrils that extends into the limbus. The natural arrangement of collagen in the chicken cornea is discussed in relation to corneal shape and the mechanical requirements of avian corneal accommodation. Equivalent data are also presented from age-matched blind chickens affected with the retinopathy, globe enlarged (rge) mutation, characterized by an abnormally thick and flat cornea. The data indicate considerable realignment and redistribution of collagen lamellae in the peripheral rge cornea. In contrast to normal chickens, no obvious tangential collagen alignment was evident in the periphery of rge corneas. In mammals, the presence of a limbal fibril annulus is believed to be important in corneal shape preservation. We postulate that corneal flattening in rge chickens may be related to biomechanical changes brought about by an alteration in collagen arrangement at the corneal periphery.

Intrastromal Injection of China Painting Ink in Corneas of Male Rabbits: Clinical and Histological Study.

PubMed

Alsmman Hassan, Alahmady Hamad; Abd Elhaliem Soliman, Nesreen Gamal-Eldeen

2016-01-01

Background. Many patients with corneal opacity or complicated cataract in blind eye ask for cosmoses. In this study we tried to investigate the staining of corneas of male rabbits by Rotring China painting ink and to study the histological changes. Method. 10 eyes of 10 male Baladi Egyptian rabbits were injected (0.1 mL) intrastromally in the cornea by the use of China painting ink (Rotring Tinta China) through insulin syringe (27-gauge needle) by single injection; clinical follow-up is for 6 months and lastly the rabbits were scarified and the stained eyes were enucleated for histological analysis. Results. Clinically the stain was stable in color and distribution in corneas with no major complications. Histological results of the stained rabbit corneas showed blackish pigmentation in the corneal stroma without any inflammatory cellular infiltration. Some fibroblast cells had pigment granules in their cytoplasm in the adjacent layers. Conclusion. Corneal staining by China painting ink is effective and safe in staining of male rabbits cornea; however further study in human corneas with longer follow-up period is advisable.

Dual interferometer for dynamic measurement of corneal topography

NASA Astrophysics Data System (ADS)

Micali, Jason D.; Greivenkamp, John E.

2016-08-01

The cornea is the anterior most surface of the eye and plays a critical role in vision. A thin fluid layer, the tear film, coats the outer surface of the cornea and serves to protect, nourish, and lubricate the cornea. At the same time, the tear film is responsible for creating a smooth continuous surface, where the majority of refraction takes place in the eye. A significant component of vision quality is determined by the shape of the cornea and stability of the tear film. A dual interferometer system for measuring the dynamic corneal topography is designed, built, verified, and qualified by testing on human subjects. The system consists of two coaligned simultaneous phase-shifting polarization-splitting Twyman-Green interferometers. The primary interferometer measures the surface of the tear film while the secondary interferometer tracks the absolute position of the cornea, which provides enough information to reconstruct the absolute shape of the cornea. The results are high-resolution and high-accuracy surface topography measurements of the in vivo tear film and cornea that are captured at standard camera frame rates.

Residual corneal stroma in big-bubble deep anterior lamellar keratoplasty: a histological study in eye-bank corneas.

PubMed

McKee, Hamish D; Irion, Luciane C D; Carley, Fiona M; Jhanji, Vishal; Brahma, Arun K

2011-10-01

To determine if residual corneal stroma remains on the recipient posterior lamella in big-bubble deep anterior lamellar keratoplasty (DALK). Pneumodissection using the big-bubble technique was carried out on eye-bank corneas mounted on an artificial anterior chamber. Samples that had a successful big-bubble formation were sent for histological evaluation to determine if any residual stroma remained on the Descemet membrane (DM). Big-bubble formation was achieved in 32 donor corneas. Two distinct types of big-bubble were seen: the bubble had either a white margin (30 corneas) or a clear margin (two corneas). The posterior lamellae of all the white margin corneas showed residual stroma on DM with a mean central thickness of 7.0 μm (range 2.6-17.4 μm). The clear margin corneas showed no residual stroma on DM. It should no longer be assumed that big-bubble DALK, where the bubble has a white margin, routinely bares DM. True baring of DM may only occur with the less commonly seen clear margin bubble.

Angiogenesis and Therapeutic Approaches to NF1 Tumors

DTIC Science & Technology

2007-04-01

corneal neovascularization model was developed. In this model, the avascularity of the cornea highly facilitates the quantification of neovascularture...wild-type corneas in avascular area. However, in the NV zone, the number of macrophage was 4.6-fold greater in Nf1þ /– corneas than wild-type corneas...GEM tumor classification because of low cellularity and no necrosis . They exceed that clas- sification, however, due to their low to moderate prolif

DOE Office of Scientific and Technical Information (OSTI.GOV)

Larin, Kirill V; Ghosn, M G

The passive diffusion of drugs through the epithelial surfaces of an eye (the most widespread method for medical treatment of various diseases) is considered. The permeability of water and drugs through rabbit cornea was measured in the isolated cornea (separate from an eye) and in the whole cornea. The permeability coefficients of water and dexamethasone were estimated by the method of optical coherence tomography (OCT). Because multiple photon scattering introduces noise and distortions to the OCT signal, measurements were performed at depths up to 500 {mu}m where most likely single scattering of light occurs in cornea. It is shown thatmore » the permeability coefficients in the isolated and whole cornea strongly differ from each other. For example, the water permeability in the isolated and whole cornea is (7.09{+-}0.12)x10{sup -5} and (1.71{+-}0.51)x10{sup -5} cm s{sup -1}, respectively. (special issue devoted to multiple radiation scattering in random media)« less

The role of crumbs genes in the vertebrate cornea.

PubMed

Beyer, Jill; Zhao, Xinping C; Yee, Richard; Khaliq, Shagufta; McMahon, Timothy T; Ying, Hongyu; Yue, Beatrice Y J T; Malicki, Jarema J

2010-09-01

To evaluate the role of crumbs genes and related epithelial polarity loci in the vertebrate cornea. The authors used histologic analysis and electron microscopy to evaluate the corneas of zebrafish mutant for a crumbs locus oko meduzy (ome) and in mutants of four other loci, nagie oko (nok), heart and soul (has), mosaic eyes (moe), and ncad (formerly glass onion), that function in the same or related genetic pathways. In parallel, they performed an evaluation of corneas in human carriers of a crumbs gene, CRB1, and mutations using topography and biomicroscopy. The expression of the CRB1 gene in the normal human cornea was examined by polymerase chain reaction (PCR) and immunohistochemical staining. The corneas of zebrafish mutants display severe abnormalities of the epithelial and stromal layers. The epithelial cells do not properly adhere to each other, and fluid-filled spaces form between them. In addition, the layering of the corneal stroma is poorly formed or absent. The corneas of human carriers of CRB1 mutations display shape deviations compared with what has been observed in normal individuals. A PCR product of the correct size was obtained from normal human corneal samples. Sequence analyses confirmed its identity to be the human CRB1 gene. Immunohistochemical staining using anti-CRB1 yielded positive brown deposits in the human cornea. crumbs genes play a role in the differentiation of the vertebrate cornea. Corneal defects associated with crumbs gene mutations are very severe in the zebrafish model and, in comparison, appear clinically less pronounced in the human eye.

Impact of the Cornea Donor Study (CDS) on Acceptance of Corneas from Older Donors

PubMed Central

Sugar, Alan; Montoya, Monty M.; Beck, Roy; Cowden, John W.; Dontchev, Mariya; Gal, Robin L.; Kollman, Craig; Malling, Jackie; Mannis, Mark J.; Tennant, Bradley

2014-01-01

Purpose Evaluate retrospectively whether findings from the Cornea Donor Study (CDS) led to changes in the transplantation of corneas from older donors. Methods United States eye banks provided complete data on donor age and placement (domestic or international) for 86,273 corneas from 1998 to 2009. The data were analyzed by 3 time periods: preceding CDS (1998–1999), during CDS (2000–2007) and after publication of CDS 5 year results (2008–2009), and separately for corneas placed within vs. outside the United States. Results For corneal tissues transplanted in the United States, the percentage of donors ≥66 years old increased from 19% before CDS to 21% during CDS and 25% after CDS (p<0.001). Corresponding median (25th-75th percentile) donor ages were 53 (39–63), 54 (41–64) and 57 (46–66), respectively (p<0.001). The opposite trend was observed for corneas distributed outside the United States with the percentage of donors ≥66 years old decreasing from 56% to 42% to 34%, respectively. Donor age trends over time varied by eye bank. Conclusions There was a modest overall increase in the donor age of corneas transplanted in the United States from 1998 to 2009, but the retrospective nature of the study limits our ability to attribute this change to the CDS. The modest increases in the donor age of corneas transplanted is a positive finding, but wider acceptance of older corneal donor tissue should be encouraged based on the five-year evidence generated by the CDS. PMID:22262218

Effect of dehydration in the UV transmittance of "in vitro" corneas

NASA Astrophysics Data System (ADS)

Lincoln, Victor A. C.; Ventura, Liliane; Faria e Sousa, Sidney J.; Mello, Marcio M.

2012-03-01

In ophthalmology the research using "in vitro" corneas are an excellent model for studies of new ophthalmologic procedures, enabling the analysis of effectiveness, performance and even safety parameters of the procedure. In this work we studied four "in vitro" human corneas preserved in OPTISOL-GS, with initial average pachymetry of 542 microns and a post-mortem average of 6 days. The corneas were preserved in OPTISOL-GS and were washed with saline solution to remove the excess the preservative medium. The corneas were placed in a device aligned with an ultraviolet source of 3mw/cm2 and an optical fiber positioned after the device near the endothelium of the cornea. The UV transmittance spectra in the region of 360-370nm were captured by the emission of UV source for 3 seconds. These spectra were captured every 5 minutes in a total of 60 minutes, resulting in 13 spectra per cornea. The measured average initial UV transmittance was 73% and after 50 minutes of dehydration there was no significant difference in the corneal teansmittance properties. However, for the last 10 minutes we have observed a decrease in the UV transmittance of 4%, probably indicated by corneal dehydration and swelling (wrinkling of the cornea tissue. The final average pachymetry was 421 microns and the UV transmittance after the 60 minutes was 69%. Therefore we can suppose that the UV transmittance of corneas "in vitro" is invariant over a period of up to 60 minutes, even with the thickness decrease, since the material that absorbs in the UV region remains intact and only water loss occurs.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cenedella, R.J.; Fleschner, C.R.

The authors developed a direct chemical approach for estimating the rate of turnover of the corneal epithelium in vivo. The method was used to examine the effects of lovastatin, a potent inhibitor of cholesterol biosynthesis, on proliferation and turnover of the epithelium. Corneal DNA was labeled by pulse injection (IP) of the rat with 3H-thymidine, and 3H-labeled DNA was recovered from peripheral and central corneas over the next 15 days. Only the epithelium became labeled, and the loss of label by cell desquamation began 3 days after injection. The loss of 3H-DNA from the cornea (peripheral plus central region) followedmore » first-order kinetics. The half-life of the disappearance was about 3 days. The peripheral cornea became more highly labeled than the central cornea and began to lose 3H-DNA before the central cornea. These observations support the possibility of a higher mitotic rate in the peripheral region and the centripetal movement of a population of peripheral epithelial cells in the normal cornea. The half-lives of the disappearance of 3H-DNA from peripheral and central corneas measured between days 5 and 15 postinjection were identical, both at 3 days. Complete turnover of the corneal epithelium would, therefore, require about 2 weeks (4-5 half-lives). Treatment of the rat with lovastatin had no obvious effects upon the proliferation or turnover of the corneal epithelium. Although lovastatin inhibited corneal 3-hydroxy-3-methylglutaryl coenzyme A reductase, the key regulatory enzyme of cholesterol synthesis, the cornea compensated by induction of this enzyme so that there was no net inhibition of cholesterol synthesis in the cornea.« less

Treatment of alkali-injured cornea by cyclosporine A-loaded electrospun nanofibers - An alternative mode of therapy.

PubMed

Cejkova, Jitka; Cejka, Cestmir; Trosan, Peter; Zajicova, Alena; Sykova, Eva; Holan, Vladimir

2016-06-01

In this study we tried to develop a new approach to suppress inflammation and neovascularization in the alkali-injured rabbit cornea. For this reason Cyclosporine A (CsA)-loaded electrospun nanofibers were transferred onto the ocular surface injured with alkali (0.25 N NaOH). Damaged corneas were divided into the following groups: untreated, treated with CsA eye drops, treated with nanofibers drug-free and treated with CsA-loaded nanofibers. Healthy rabbit corneas served as controls. Drug-free nanofibers and CsA-loaded nanofibers were transferred onto the damaged corneal surface immediately after the injury and sutured to conjunctiva. On day five after the injury the nanofibers were removed. The animals from all groups were sacrificed on day twelve after the injury. The extent of the inflammatory reaction and corneal healing were examined macroscopically, immunohistochemically and biochemically. The central corneal thickness was measured using an ultrasonic pachymeter. When compared with untreated injured corneas, injured corneas treated with drug-free nanofibers or injured corneas treated with CsA eye drops, the number of CD3-positive cells (T lymphocytes) and the production of pro-inflammatory cytokines were strongly reduced in corneas treated with CsA-loaded nanofibers, which was associated with the significantly decreased expression of matrix metalloproteinase 9, inducible nitric oxide synthase, vascular endothelial growth factor and active caspase-3. CsA-loaded nanofibers effectively suppressed corneal inflammation and corneal neovascularization. Central corneal thickness restored to levels before injury only in corneas treated with CsA-loaded nanofibers. Corneal transparency was highly restored in these corneas. It is suggested that the beneficial effect of CsA-loaded nanofibers was associated with the continuous release of CsA from nanofibers and continuous affection of damaged cornea by CsA. The suture of nanofibers to conjunctiva and the closed eyes contributed to beneficial corneal healing. This is in contrast to CsA eye drops, which are quickly washed from the ocular surface and the contact of CsA with the damaged cornea was limited. In conclusion, the approach with CsA-loaded nanofibers could represent an effective alternative mode of therapy for corneal chemical burns. Copyright © 2016 Elsevier Ltd. All rights reserved.

Keratoconus corneal architecture after riboflavin/ultraviolet A cross-linking: Ultrastructural studies

PubMed Central

Almubrad, Turki; Paladini, Iacopo; Mencucci, Rita

2013-01-01

Purpose Study to investigate the effects of collagen cross-linking on the ultrastructural organization of the corneal stroma in the human keratoconus cornea (KC). Methods Three normal, three keratoconus (KC1, KC2, KC3), and three cross-linked keratoconus (CXL1, CXL2, CXL3) corneas were analyzed. The KC corneas were treated with a riboflavin-ultraviolet A (UVA) treatment (CXL) method described by Wollensak et al. Penetrating keratoplasty (PKP) was performed 6 months after treatment. All samples were processed for electron microscopy. Results The riboflavin-UVA-treated CXL corneal stroma showed interlacing lamellae in the anterior stroma followed by well-organized parallel running lamellae. The lamellae contained uniformly distributed collagen fibrils (CFs) decorated with normal proteoglycans (PGs). The CF diameter and interfibrillar spacing in the CXL cornea were significantly increased compared to those in the KC cornea. The PG area in the CXL corneas were significantly smaller than the PGs in the KC cornea. The epithelium and Bowman’s layer were also normal. On rare occasions, a thick basement membrane and collagenous pannus were also observed. Conclusions Corneal cross-linking leads to modifications of the cornea stroma. The KC corneal structure showed a modification in the CF diameter, interfibrillar spacing, and PG area. This resulted in a more uniform distribution of collagen fibrils, a key feature for corneal transparency. PMID:23878503

The Naïve Murine Cornea as a Model System to Identify Novel Endogenous Regulators of Lymphangiogenesis: TRAIL and rtPA.

PubMed

Regenfuß, Birgit; Dreisow, Marie-Luise; Hos, Deniz; Masli, Sharmila; Bock, Felix; Cursiefen, Claus

2015-06-01

In the murine cornea, which is an established model for analyzing pathologic lymphatic vessel growth, phenotypic heterogeneity of the endogenous lymphatic vessels in the limbus of the cornea was previously described. In this study, the cornea of BALB/c, C57BL/6, and FVB mice with different limbal lymphangiogenic phenotypes was analyzed to identify novel candidates potentially influencing lymphatic vessel growth. Pathway specific expression analysis of the cornea was performed to identify novel candidate genes. Corneal protein expression of the respective candidates was analyzed by fluorescent immunohistochemistry. The effect of the candidates on proliferation of human dermal lymphatic endothelial cells (HDLECs) was analyzed by BrdU proliferation ELISA. Thirteen genes were differentially regulated in corneas of mouse strains with more endogenous limbal lymphatic vessels (high-lymphangiogenic) (C57BL/6) compared to mouse strains with less endogenous limbal lymphatic vessels (low-lymphangiogenic) (BALB/c, FVB). Two candidates, Tumor necrosis factor (ligand) superfamily member 10 (Tnfsf10/Trail) and Plasminogen activator, tissue (Plat/tPA) were expressed in the cornea of BALB/c and C57BL/6 mice on the protein level. In vitro, Trail and recombinant tPA inhibited the proliferation of human dermal lymphatic endothelial cells. Molecular analysis of the naive cornea in mouse strains with different limbal lymphatic phenotypes is a valuable model to identify novel endogenous regulators of lymphangiogenesis.

The effect of actinoquinol with hyaluronic acid in eye drops on the optical properties and oxidative damage of the rabbit cornea irradiated with UVB rays.

PubMed

Čejka, Čestmír; Luyckx, Jacques; Ardan, Taras; Pláteník, Jan; Širc, Jakub; Michálek, Jiří; Čejková, Jitka

2010-01-01

Irradiation of the cornea with UVB rays leads to its oxidative damage, swelling and increased light absorption. We investigated changes in the corneal optics (evaluated by changes of corneal hydration and light absorption) and microscopical disturbances of corneas irradiated with UVB rays as influenced by eye drops containing actinoquinol with hyaluronic acid. Rabbit corneas were irradiated with a daily dose of 0.5 or 1.01 J cm(-2) of UVB rays (312 nm) for 4 days. During irradiation, the eye drops were applied on the right eye and buffered saline (or hyaluronic acid) on the left eye. On day 5 the rabbits were sacrificed and the corneas examined spectrophotometrically for light absorption. The corneal thickness (hydration) was measured using a pachymeter. Corneas of some other rabbits were examined immunohistochemically. After buffered saline treatment UVB rays evoked changes in the corneal optics and induced oxidative damage of the corneas. After actinoquinol-hyaluronic acid application, these changes were diminished. Hyaluronic acid alone was less effective. In conclusion, actinoquinol-hyaluronic acid eye drops decreased changes in corneal optics and suppressed oxidative damage in the UVB-irradiated cornea. However, the effective corneal protection by these eye drops was limited to the lower UVB dose. © 2010 The Authors. Journal Compilation. The American Society of Photobiology.

Expression analysis of MDR1, BCRP and MRP3 transporter proteins in different in vitro and ex vivo cornea models for drug absorption studies.

PubMed

Verstraelen, Jessica; Reichl, Stephan

2013-01-30

Ocular drug absorption studies are required for the development of new drugs or drug delivery systems for eye treatment. Such preclinical investigations on transcorneal drug absorption are performed ex vivo with the excised corneas of experimental animals or in vitro using corneal cell culture models. The data currently available on the expression of ABC transporter proteins in corneal tissue is limited or contradictory. This study describes, for the first time, the comparison of the expression of ABC transporters, in particular, MDR1, BCRP and MRP3, between human cornea cell culture models and the most commonly used ex vivo models, namely, rabbit and porcine corneas, conducted in the same laboratory. The expression levels and functionality were determined by means of PCR, western blot, immunohistochemistry and bidirectional permeation studies using specific substrates and inhibitors. The results clearly indicate species-dependent expression of the studied efflux transporters. In the rabbit cornea, the expression and activity of MDR1 transporter was confirmed, whereas human cell culture models and porcine corneas did not show MDR1 expression. However, human cornea models possessed MRP3 and BCRP expression, whereas no functional expression was found in rabbit and porcine corneas. Therefore, the translation of transcorneal permeation data from animal experiments to humans should be performed with caution. Copyright © 2012 Elsevier B.V. All rights reserved.

Hydration and transparency of the rabbit cornea irradiated with UVB-doses of 0.25 J/cm(2) and 0.5 J/cm(2) compared with equivalent UVB radiation exposure reaching the human cornea from sunlight.

PubMed

Cejka, Cestmír; Ardan, Taras; Sirc, Jakub; Michálek, Jiří; Beneš, Jiří; Brůnová, Blanka; Rosina, Jozef

2011-07-01

Exposure of the cornea to UV radiation from sunlight evokes intraocular inflammation, photokeratitis. Photokeratitis is caused by UVB radiation. It is accompanied by changes of corneal hydration and light absorption. The aim of this study was to examine the effect of two UVB doses on corneal optics in rabbits and to compare these UVB doses with the equivalent exposure of UVB radiation reaching the human cornea from sunlight. Rabbit corneas were irradiated with a daily UVB dose of 0.25 J/cm(2) or 0.5 J/cm(2) for 4 days. One day after finishing the irradiations the rabbits were sacrificed and corneal light absorption measured using our spectrophotometrical method. Corneal hydration was examined using an ultrasonic Pachymeter every experimental day before the irradiation procedure and the last day before sacrificing the animals. Changes in corneal optics appeared after the repeated exposure of the cornea to a UVB dose of 0.25 J/ cm(2) and massively increased after the repeated exposure of the cornea to a UVB dose of 0.5 J/cm(2). The first significant changes in corneal hydration appeared after a single exposure of the cornea to a UVB dose of 0.25 J/cm(2). Changes in corneal hydration appeared after the exposure of the rabbit cornea to a single UVB dose equivalent to 2.6 hours of solar UVB radiation reaching the human cornea, as measured by UVB sensors embedded in the eyes of mannequin heads facing the sun on a beach at noon in July. Repeated exposure of the rabbit cornea to the same UVB dose evoked profound changes in corneal optics. Although comparison of experimental and outdoor conditions are only approximate, the results in rabbits point to the danger for the human eye from UVB radiation when short stays in sunlight are repeated for several consecutive days without UV protection.

Stromal fibroblasts are associated with collagen IV in scar tissues of alkali-burned and lacerated corneas.

PubMed

Ishizaki, M; Shimoda, M; Wakamatsu, K; Ogro, T; Yamanaka, N; Kao, C W; Kao, W W

1997-04-01

Corneal wound healing frequently leads to the formation of opaque scar tissue. We examined whether stromal fibroblastic cells of injured corneas express collagen IV and contributes to the formation of a basal lamina-like structure. Rabbits were anesthetized, and central corneal alkali burn (8 mm in diameter; 1 M NaOH, 1 min) or laceration (8 mm long) were produced. The injured corneas, which had healed for 1, 7, 21 and 45 days, were subjected to histological and immunohistochemical studies with goat anti-collagen IV antibodies, using light and electron microscopy, and in situ hybridization with an antisense digoxigenin-labeled riboprobe of collagen alpha 1(IV) mRNA. For comparison, twenty-day-old fetal corneas were subjected to immunohistochemical study and transmission electron microscopy (TEM). TEM examinations revealed that the stromal collagenous matrix was organized in orthogonal lamellae during corneal development, whereas that of alkali-burned cornea, which had healed for 3 weeks, was disorganized. The stroma of twenty-day-old fetal cornea was not labeled by the anti-collagen IV antibodies. In contrast, one week after injury, specific collagen IV immunostaining was detected in the injured stroma. As the healing proceeded (21-45 days), the antibodies reacted with fibroblastic cells and the extracellular matrix of scar tissues located in the anterior portion of alkali-burned corneas, as well as the posterior portion of lacerated corneas. The middle portion of the stromal tissues was weakly labeled by the anti-collagen IV antibodies with the exception of the blood vessel wall. Immuno-electron microscopic study showed that collagen IV and fibronectin were closely associated with the fibroblastic cells. In situ hybridization demonstrated that epithelial and endothelial cells and fibroblastic cells in the wounded corneal stroma and retro-corneal membrane expressed alpha 1(IV) mRNA, whereas in normal corneas the expression of alpha 1(IV) mRNA was limited to epithelial and endothelial cells. The enhanced expression of collagen IV by the fibroblastic cells in the stroma of injured corneas is consistent with the notion that they may contribute to the formation of basal lamina-like structures in injured corneas.

Detection of Differentially Expressed Wound-Healing–Related Glycogenes in Galectin-3–Deficient Mice

PubMed Central

Saravanan, Chandrassegar; Cao, Zhiyi; Head, Steven R.; Panjwani, Noorjahan

2010-01-01

Purpose A prior study showed that exogenous galectin-3 (Gal-3) stimulates re-epithelialization of corneal wounds in wild-type (Gal-3+/+) mice but, surprisingly, not in galectin-3–deficient (Gal-3−/−) mice. In an effort to understand why the injured corneas of Gal-3−/− mice are unresponsive to exogenous Gal-3, the present study was designed to determine whether genes encoding the enzymes that regulate the synthesis of glycan ligands of Gal-3 are differentially expressed in Gal-3−/− corneas compared with the Gal-3+/+ corneas. Methods Glycogene microarray technology was used to identify differentially expressed glycosyltransferases in healing Gal-3+/+ and Gal-3−/− corneas. Results Of ~2000 glycogenes on the array, the expression of 8 was upregulated and that of 14 was downregulated more than 1.3-fold in healing Gal-3−/− corneas. A galactosyltransferase, β3GalT5, which has the ability to synthesize Gal-3 ligands was markedly downregulated in healing Gal-3−/− corneas. The genes for polypeptide galactosaminyltransferases (ppGalNAcT-3 and -7) that are known to initiate O-linked glycosylation and N-aspartyl-β-glucosaminidase, which participates in the removal of N-glycans, were found to be upregulated in healing Gal-3−/− corneas. Microarray data were validated by qRT-PCR. Conclusions Based on the known functions of the differentially expressed glycogenes, it appears that the glycan structures on glycoproteins and glycolipids, synthesized as a result of the differential glycogene expression pattern in healing Gal-3−/− corneas may lead to the downregulation of specific counterreceptors for Gal-3. This may explain, at least in part, why, unlike healing Gal-3+/+ corneas, the healing Gal-3−/− corneas are unresponsive to the stimulatory effect of exogenous Gal-3 on re-epithelialization of corneal wounds. PMID:19643959

Simulation analysis of the transparency of cornea and sclera

NASA Astrophysics Data System (ADS)

Yang, Chih-Yao; Tseng, Snow H.

2017-02-01

Both consist of collagen fibrils, sclera is opaque whereas cornea is transparent for optical wavelengths. By employing the pseudospectral time-domain (PSTD) simulation technique, we model light impinging upon cornea and sclera, respectively. To analyze the scattering characteristics of light, the cornea and sclera are modeled by different sizes and arrangements of the non-absorbing collagen fibrils. Various factors are analyzed, including the wavelength of incident light, the thickness of the scattering media, position of the collagen fibrils, size distribution of the fibrils.

Serum response factor: positive and negative regulation of an epithelial gene expression network in the destrin mutant cornea

PubMed Central

Kawakami-Schulz, Sharolyn V.; Verdoni, Angela M.; Sattler, Shannon G.; Jessen, Erik; Kao, Winston W.-Y.; Ikeda, Akihiro

2014-01-01

Increased angiogenesis, inflammation, and proliferation are hallmarks of diseased tissues, and in vivo models of these disease phenotypes can provide insight into disease pathology. Dstncorn1 mice, deficient for the actin depolymerizing factor destrin (DSTN), display an increase of serum response factor (SRF) that results in epithelial hyperproliferation, inflammation, and neovascularization in the cornea. Previous work demonstrated that conditional ablation of Srf from the corneal epithelium of Dstncorn1 mice returns the cornea to a wild-type (WT) like state. This result implicated SRF as a major regulator of genes that contributes to abnormal phenotypes in Dstncorn1 cornea. The purpose of this study is to identify gene networks that are affected by increased expression of Srf in the Dstncorn1 cornea. Microarray analysis led to characterization of gene expression changes that occur when conditional knockout of Srf rescues mutant phenotypes in the cornea of Dstncorn1 mice. Comparison of gene expression values from WT, Dstncorn1 mutant, and Dstncorn1 rescued cornea identified >400 differentially expressed genes that are downstream from SRF. Srf ablation had a significant effect on genes associated with epithelial cell-cell junctions and regulation of actin dynamics. The majority of genes affected by SRF are downregulated in the Dstncorn1 mutant cornea, suggesting that increased SRF negatively affects transcription of SRF gene targets. ChIP-seq analysis on Dstncorn1 mutant and WT tissue revealed that, despite being present in higher abundance, SRF binding is significantly decreased in the Dstncorn1 mutant cornea. This study uses a unique model combining genetic and genomic approaches to identify genes that are regulated by SRF. These findings expand current understanding of the role of SRF in both normal and abnormal tissue homeostasis. PMID:24550211

Material Properties from Air Puff Corneal Deformation by Numerical Simulations on Model Corneas.

PubMed

Bekesi, Nandor; Dorronsoro, Carlos; de la Hoz, Andrés; Marcos, Susana

2016-01-01

To validate a new method for reconstructing corneal biomechanical properties from air puff corneal deformation images using hydrogel polymer model corneas and porcine corneas. Air puff deformation imaging was performed on model eyes with artificial corneas made out of three different hydrogel materials with three different thicknesses and on porcine eyes, at constant intraocular pressure of 15 mmHg. The cornea air puff deformation was modeled using finite elements, and hyperelastic material parameters were determined through inverse modeling, minimizing the difference between the simulated and the measured central deformation amplitude and central-peripheral deformation ratio parameters. Uniaxial tensile tests were performed on the model cornea materials as well as on corneal strips, and the results were compared to stress-strain simulations assuming the reconstructed material parameters. The measured and simulated spatial and temporal profiles of the air puff deformation tests were in good agreement (< 7% average discrepancy). The simulated stress-strain curves of the studied hydrogel corneal materials fitted well the experimental stress-strain curves from uniaxial extensiometry, particularly in the 0-0.4 range. Equivalent Young´s moduli of the reconstructed material properties from air-puff were 0.31, 0.58 and 0.48 MPa for the three polymer materials respectively which differed < 1% from those obtained from extensiometry. The simulations of the same material but different thickness resulted in similar reconstructed material properties. The air-puff reconstructed average equivalent Young´s modulus of the porcine corneas was 1.3 MPa, within 18% of that obtained from extensiometry. Air puff corneal deformation imaging with inverse finite element modeling can retrieve material properties of model hydrogel polymer corneas and real corneas, which are in good correspondence with those obtained from uniaxial extensiometry, suggesting that this is a promising technique to retrieve quantitative corneal biomechanical properties.

Comparison of endothelial cell density of organ cultured corneas with cornea donor study.

PubMed

Campolmi, Nelly; He, Zhiguo; Acquart, Sophie; Trone, Marie-Caroline; Bernard, Aurélien; Gauthier, Anne-Sophie; Garraud, Olivier; Forest, Fabien; Péocʼh, Michel; Gain, Philippe; Thuret, Gilles

2014-06-01

Determination of the endothelial cell density (ECD) by eye banks is paramount in donor cornea qualification. Unbiased measurement avoids wastage and grafts with an increased risk of premature failure. Internal calibration of the counting method is essential, but external validation would add an extra stage in the assessment of reliability. In this respect, data published by the multicenter Cornea Donor Study (CDS) in 2005 is a reference. The aim of the study was to compare ECD determined within a single eye bank, which uses calibrated image analysis software designed for transmitted light microscopy images of organ cultured corneas, with the CDS data determined on specular microscopy images of corneas stored at 4°C. ECD of consecutive corneas retrieved between 2005 and 2013 was determined after exposure to 0.9% NaCl. More than 300 ECs were counted on 3 fields of the central 8 mm. Endothelial cell boundaries were automatically drawn and verified by a skilled technician who performed all necessary corrections. Three thousand fifty-two corneas were analyzed, of which 48.5% donors were >75 years (CDS upper age limit). Between 10 and 75 years, the ECD varied according to donor age exactly in the same manner as in the CDS, but were consistently higher of 100 ± 25 cells per square millimeter (P < 0.001). ECD determined by a computer-aided method from transmitted light microscopy images compares favorably with the American CDS reference series. The slight systematic difference on either side of the Atlantic Ocean could be due to (1) differences in counting principles and/or (2) higher shrinkage of the cornea caused by stromal edema in organ culture.

Corneal avascularity is due to soluble VEGF receptor-1

PubMed Central

Ambati, Balamurali K.; Nozaki, Miho; Singh, Nirbhai; Takeda, Atsunobu; Jani, Pooja D.; Suthar, Tushar; Albuquerque, Romulo J. C.; Richter, Elizabeth; Sakurai, Eiji; Newcomb, Michael T.; Kleinman, Mark E.; Caldwell, Ruth B.; Lin, Qing; Ogura, Yuichiro; Orecchia, Angela; Samuelson, Don A.; Agnew, Dalen W.; Leger, Judy St.; Green, W. Richard; Mahasreshti, Parameshwar J.; Curiel, David T.; Kwan, Donna; Marsh, Helene; Ikeda, Sakae; Leiper, Lucy J.; Collinson, J. Martin; Bogdanovich, Sasha; Khurana, Tejvir S.; Shibuya, Masabumi; Baldwin, Megan E.; Ferrara, Napoleone; Gerber, Hans-Peter; Falco, Sandro De; Witta, Jassir; Baffi, Judit Z.; Raisler, Brian J.; Ambati, Jayakrishna

2009-01-01

Corneal avascularity—the absence of blood vessels in the cornea—is required for optical clarity and optimal vision, and has led to the cornea being widely used for validating pro- and anti-angiogenic therapeutic strategies for many disorders1-4. But the molecular underpinnings of the avascular phenotype have until now remained obscure5-10 and are all the more remarkable given the presence in the cornea of vascular endothelial growth factor (VEGF)-A, a potent stimulator of angiogenesis, and the proximity of the cornea to vascularized tissues. Here we show that the cornea expresses soluble VEGF receptor-1 (sVEGFR-1; also known as sflt-1) and that suppression of this endogenous VEGF-A trap11 by neutralizing antibodies, RNA interference or Cre-lox-mediated gene disruption abolishes corneal avascularity in mice. The spontaneously vascularized corneas of corn1 and Pax6+/− mice12,13 and Pax6+/− patients with aniridia14 are deficient in sflt-1, and recombinant sflt-1 administration restores corneal avascularity in corn1 and Pax6+/− mice. Manatees, the only known creatures uniformly to have vascularized corneas15, do not express sflt-1, whereas the avascular corneas of dugongs, also members of the order Sirenia, elephants, the closest extant terrestrial phylogenetic relatives of manatees, and other marine mammals (dolphins and whales) contain sflt-1, indicating that it has a crucial, evolutionarily conserved role. The recognition that sflt-1 is essential for preserving the avascular ambit of the cornea can rationally guide its use as a platform for angiogenic modulators, supports its use in treating neovascular diseases, and might provide insight into the immunological privilege of the cornea. PMID:17051153

DOE Office of Scientific and Technical Information (OSTI.GOV)

Quantock,A.; Boote, C.; Young, R.

In the cornea of the eye light transmission is facilitated by the regular arrangement and uniform diameter of collagen fibrils that constitute the bulk of the extracellular corneal matrix. Matrix architecture, in turn, is believed to be governed by interactions between collagen fibrils and proteoglycan molecules modified with sulfated glycosaminoglycan side chains. Here, we outline the contribution made by small-angle X-ray scattering studies of the cornea in understanding the role of sulfated glycosaminoglycans in the control of collagen architecture in cornea, and present new depth-profiled microbeam data from swollen human eye-bank corneas that indicate no significant change in collagen fibrilmore » diameter throughout the tissue, but a lower collagen interfibrillar spacing in the anterior-most stromal regions compared with the ultrastructure of the deeper cornea.« less

[Results of an internet-based survey amongst members of the German Ophthalmological Society concerning postmortem cornea donation].

PubMed

Uhlig, C E; Promesberger, J; Hirschfeld, G; Koch, R; Reinhard, T; Seitz, B

2012-12-01

Analysis of willingness for postmortem cornea donation by professionals in ophthalmology and their motives in favor of or against donation. 3887 members of the German Ophthalmological Society received an anonymous questionnaire concerning sociodemographic background, physical health, experiences with organ explantation and their former engagement and motives concerning organ and cornea donation. 722 of the questionnaires were partially and 533 completely answered with an average willingness for cornea donation of 79.4%. Significant parameters for cornea donation were gender, former experience with organ explantation, ophthalmological health and fear of false diagnosis of brain death, worse medical treatment or organ commercialization. Of the participants 53.9% suggested the internet as a favorite source of information in this matter. The factors which had a significant impact on cornea donation in this survey seem to be mainly a result of insufficient information. Detailed information regarding this topic should preferentially be presented on internet pages of professional societies and could probably increase donation approval of DOG members.

Portable light transmission measuring system for preserved corneas.

PubMed

Ventura, Liliane; Jesus, Gabriel Torres de; Oliveira, Gunter Camilo Dablas de; Sousa, Sidney J F

2005-12-22

The authors have developed a small portable device for the objective measurement of the transparency of corneas stored in preservative medium, for use by eye banks in evaluation prior to transplantation. The optical system consists of a white light, lenses, and pinholes that collimate the white light beams and illuminate the cornea in its preservative medium, and an optical filter (400-700 nm) that selects the range of the wavelength of interest. A sensor detects the light that passes through the cornea, and the average corneal transparency is displayed. In order to obtain only the tissue transparency, an electronic circuit was built to detect a baseline input of the preservative medium prior to the measurement of corneal transparency. The operation of the system involves three steps: adjusting the "0 %" transmittance of the instrument, determining the "100 %" transmittance of the system, and finally measuring the transparency of the preserved cornea inside the storage medium. Fifty selected corneas were evaluated. Each cornea was submitted to three evaluation methods: subjective classification of transparency through a slit lamp, quantification of the transmittance of light using a corneal spectrophotometer previously developed, and measurement of transparency with the portable device. By comparing the three methods and using the expertise of eye bank trained personnel, a table for quantifying corneal transparency with the new device has been developed. The correlation factor between the corneal spectrophotometer and the new device is 0,99813, leading to a system that is able to standardize transparency measurements of preserved corneas, which is currently done subjectively.

Effects of the holmium laser on the human cornea: a preliminary study

NASA Astrophysics Data System (ADS)

Mueller, Linda J.; Tassignon, Marie J.; Trau, Rene; Pels, Liesbeth; Vrensen, Gijs F.

1996-12-01

Treatment of peripheral post-mortem human corneas with the Holmium laser in a ring pattern resulted in opaque spots. One pair of treated eyes was immediately processed for light and electron microscopy and three other treated eyes were preserved for 4 days in medium in order to compare direct and short-term effects of the Holmium laser. Cross as well as frontal light microscopical sections of all eyes revealed interconnecting bands between the spots. At the ultrastructural level the anterior corneal tissue within these spots was characterized by coagulation of cells and collagen and shoed either a dramatic distorting effect on the epithelium in the eyes processed immediately or a single layer of flattened multi-nucleolated epithelial cells having more than one nucleolus per nucleus in the eyes stored in medium. Furthermore, the spots showed disturbed Bowman's layer, destroyed keratocytes and collagen fibrils which were either coagulated or organized chaotically. The interconnecting bands contained alternating normal and coagulated collagen fibers. The rest of the cornea outside the spots had a normal appearance. In corneas stored in medium, both keratocytes and epithelial cells over the entire cornea exhibited accumulations of cytoplasmic fibrils and glycogen particles. These phenomena were not observed in non-preserved corneas, suggesting that the differences are due to preservation and not due to the laser treatment. It is concluded that morphological changes occur mainly in the treated peripheral cornea whereas the central untreated cornea remains unaffected,indicating that the Holmium laser is a reliable instrument to treat hypermetropic patients.

Multiphoton Imaging of Rabbit Cornea Treated with Mitomycin C after Photorefractive Keratectomy

NASA Astrophysics Data System (ADS)

Hsueh, Chiu-Mei; Lo, Wen; Wang, Tsung-Jen; Hu, Fung-Rong; Dong, Chen-Yuan

2007-07-01

In this work we use multiphoton microscopy to observe the post surgery structure variation of rabbit cornea after photorefractive keratectomy (PRK). In addition, we added mitomycin C (MMC) to the post surgery rabbit cornea in order to investigate the effect of MMC treatment on the postoperative regeneration.

Anatomy and physiology of the cornea.

PubMed

DelMonte, Derek W; Kim, Terry

2011-03-01

The importance of the cornea to the ocular structure and visual system is often overlooked because of the cornea's unassuming transparent nature. The cornea lacks the neurobiological sophistication of the retina and the dynamic movement of the lens; yet, without its clarity, the eye would not be able to perform its necessary functions. The complexity of structure and function necessary to maintain such elegant simplicity is the wonder that draws us to one of the most important components of our visual system. Copyright © 2011 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

Combined microwave heating and surface cooling of the cornea.

PubMed

Trembly, B S; Keates, R H

1991-01-01

We investigated a nonsurgical means of reshaping the cornea to correct hyperopia, keratoconus, or myopia. The object was to heat the central stroma of the cornea to the shrinkage temperature of collagen, 55-58 degrees C. The heating device was an open-ended, coaxial, near-field applicator driven at 2450 MHz; it incorporates cooling of the cornea surface by flow of saline. We investigated the system theoretically by computing the 2-D, axisymmetric temperature distribution with the finite element method. We investigated the system experimentally by heating excised steer corneas. Histology showed the system could shrink the stroma to a depth of 0.6 mm while sparing the epithelium in 75% of cases; the diameter of shrinkage was 1.3 mm. Theory predicted a significantly deeper and narrower region of shrinkage than was observed.

Preparation of pre-cut corneas from fresh donated whole globes for Descemet's stripping automated keratoplasty: 3-year results at the Central Eye Bank of Iran.

PubMed

Kanavi, Mozhgan Rezaei; Javadi, Mohammad Ali; Javadi, Fatemeh; Chamani, Tahereh

2014-09-01

To describe the technique and the results of the preparation of pre-cut corneas for Descemet's stripping automated endothelial keratoplasty (DSAEK) during a 3-year period at the Central Eye Bank of Iran (CEBI). The method of preparation of pre-cut corneas from donated whole globes at the CEBI is described and the frequency and percentage of pre-cut corneas prepared for DSAEK, between April 2009 and March 2012, are specified. Moreover, post-operative reports are reviewed for any complaints about using pre-cut tissues for DSAEK. Out of the 1,518 donated whole globes appropriate for DSAEK, 1,478 (97.4 %) pre-cut corneas were successfully prepared. The method of preparation failed in 40 (2.6 %) cases. Based on the eye bank post-operative reports, thickness of pre-cut tissues for DSAEK was deemed unacceptable in only 6 (0.4 %) cases prior to surgery; five of these were too thick and one was too thin. Preparation of pre-cut corneas, for DSAEK from donated whole globes, in the CEBI is a safe and easy method, with very good preservation of endothelial cells after the preparation of the pre-cut corneas and reduced risks from corneal manipulation.

Absence of Vsx1 expression in the normal and damaged mouse cornea

PubMed Central

Chow, Robert L.

2011-01-01

Purpose To examine the expression of visual system homeobox 1 (Vsx1) in the mouse cornea and its potential role in the corneal wound response pathway. Methods Expression of Vsx1 was examined by quantitative reverse-transcription PCR (qRT–PCR) in corneal tissue from developing and adult mice and from mice that had undergone alkali-burn corneal wounding. Immunolabeling and Vsx1 knock-in reporter gene expression in wild type and Vsx1 null-mice were used to confirm the qRT–PCR data. Results Using qRT–PCR, Vsx1 expression was not detected in either the postnatal or adult mouse cornea or in corneas following wounding. This qRT–PCR data was supported by the absence of specific Vsx1 immunolabeling and Vsx1 knock-in reporter expression in untreated and wounded corneas. Conclusions In mice, Vsx1 mRNA, protein or reporter gene expression is not detected in the normal or damaged cornea. These results make it uncertain what role VSX1/Vsx1 plays in corneal biology. Future experiments examining the pathogenicity of VSX1 mutations associated with corneal dystrophy are required to rule out species differences and possible non-cell autonomous roles for VSX1 in the cornea. PMID:21437200

Property-based design: optimization and characterization of polyvinyl alcohol (PVA) hydrogel and PVA-matrix composite for artificial cornea.

PubMed

Jiang, Hong; Zuo, Yi; Zhang, Li; Li, Jidong; Zhang, Aiming; Li, Yubao; Yang, Xiaochao

2014-03-01

Each approach for artificial cornea design is toward the same goal: to develop a material that best mimics the important properties of natural cornea. Accordingly, the selection and optimization of corneal substitute should be based on their physicochemical properties. In this study, three types of polyvinyl alcohol (PVA) hydrogels with different polymerization degree (PVA1799, PVA2499 and PVA2699) were prepared by freeze-thawing techniques. After characterization in terms of transparency, water content, water contact angle, mechanical property, root-mean-square roughness and protein adsorption behavior, the optimized PVA2499 hydrogel with similar properties of natural cornea was selected as a matrix material for artificial cornea. Based on this, a biomimetic artificial cornea was fabricated with core-and-skirt structure: a transparent PVA hydrogel core, surrounding by a ringed PVA-matrix composite skirt that composed of graphite, Fe-doped nano hydroxyapatite (n-Fe-HA) and PVA hydrogel. Different ratio of graphite/n-Fe-HA can tune the skirt color from dark brown to light brown, which well simulates the iris color of Oriental eyes. Moreover, morphologic and mechanical examination showed that an integrated core-and-skirt artificial cornea was formed from an interpenetrating polymer network, no phase separation appeared on the interface between the core and the skirt.

Synthetic Neurotensin Analogues Are Nontoxic Analgesics for the Rabbit Cornea

PubMed Central

Kim, Charles; Barbut, Denise; Heinemann, Murk H.; Pasternak, Gavril; Rosenblatt, Mark I.

2014-01-01

Purpose. To characterize the analgesic potency and toxicity of topical synthetic neurotensin analogues, and localize neurotensin receptors in the cornea and trigeminal ganglion. Methods. Cochet-Bonnet esthesiometry was performed on the rabbit cornea to test the analgesic dose response and duration of effect for two synthetic neurotensin analogues: NT71 and NT72. Receptors for neurotensin were localized in the murine cornea and trigeminal ganglion using quantitative PCR (qPCR), Western blotting, and immunohistochemistry. In vitro toxicity of NT71, NT72, and sodium channel blockers was evaluated using cytotoxicity, single-cell migration, and scratch closure assays performed on rabbit corneal epithelial cells. In vivo toxicity of these agents was assessed using a rabbit laser phototherapeutic keratectomy (PTK) model and histology. Results. NT71 and NT72 induced potent analgesic effects on the rabbit cornea at concentrations between 1.0 and 2.5 mg/mL, lasting up to 180 minutes. A site-specific distribution of neurotensin receptors was observed in the murine cornea and trigeminal ganglion. NT71 and NT72 did not cause any significant in vitro or in vivo toxicity, in contrast to sodium channel blockers. Conclusions. Synthetic neurotensin analogues are potent analgesics that avoid the toxicities associated with established topical analgesic agents. Receptors for neurotensin are present in both the cornea and trigeminal ganglion. PMID:24825106

Dendritic cells in the cornea during Herpes simplex viral infection and inflammation.

PubMed

Kwon, Min S; Carnt, Nicole A; Truong, Naomi R; Pattamatta, Ushasree; White, Andrew J; Samarawickrama, Chameen; Cunningham, Anthony L

Herpes simplex keratitis is commonly caused by Herpes simplex virus type 1, which primarily infects eyelids, corneas, or conjunctiva. Herpes simplex virus type 1-through sophisticated interactions with dendritic cells (DCs), a type of antigen-presenting cell)-initiates proinflammatory responses in the cornea. Corneas were once thought to be an immune-privileged region; however, with the recent discovery of DCs that reside in the cornea, this long-held conjecture has been overturned. Therefore, evaluating the clinical, preclinical, and cell-based studies that investigate the roles of DCs in corneas infected with Herpes simplex virus is critical. With in vivo confocal microscopy, animal models, and cell culture experiments, we may further the understanding of the sophisticated interactions of Herpes simplex virus with DCs in the cornea and the molecular mechanism associated with it. It has been shown that specific differentiation of DCs using immunohistochemistry, flow cytometry, and polymerase chain reaction analysis in both human and mice tissues and viral tissue infections are integral to increasing understanding. As for in vivo confocal microscopy, it holds promise as it is the least invasive and a real-time investigation. These tools will facilitate the discovery of various targets to develop new treatments. Copyright © 2017 Elsevier Inc. All rights reserved.

Numerical and clinical investigation on the material model of the cornea in Corvis tonometry tests: differentiation between hyperelasticity and viscoelasticity

NASA Astrophysics Data System (ADS)

Jannesari, Mohammad; Mosaddegh, Peiman; Kadkhodaei, Mahmoud; Kasprzak, Henryk; Jabbarvand Behrouz, Mahmoud

2018-05-01

Non-contact tonometers, including ORA and Corvis ST, are not only used to estimate intraocular pressure (IOP) in clinical surveys but are also utilized to evaluate biomechanical properties of the cornea or anterior eye. However, for the cornea a realistic material model is still a controversial issue, and the main goal of the present study is to make this clearer. To this aim, the corneal biomechanical response is modeled by using a four-element linear viscoelastic model, which is characterized by in-vivo clinical data from Corvis ST tonometer. IOP tonometry tests on 5 normal and 5 keratoconic cases are accomplished by Corvis ST tonometer. Images from cornea deformation due to applied air jet are acquired from Corvis ST and are converted to the corneal deformation profiles by image processing techniques. By excluding the eye globe rigid body motion (retraction) from the total eye displacement, pure deformation of the cornea is obtained and used to calculate the required material properties. By calculating retardation time, contribution of the material viscosity during the test is estimated. The results show that viscosity effects do not substantially contribute to the cornea response during dynamic tests for both normal and keratoconic corneas. Indeed, the viscous effect comes from the eye globe rigid body motion.

The structural response of the cornea to changes in stromal hydration

PubMed Central

White, Tomas; Boote, Craig; Kamma-Lorger, Christina S.; Bell, James; Sorenson, Thomas; Terrill, Nick; Shebanova, Olga; Meek, Keith M.

2017-01-01

The primary aim of this study was to quantify the relationship between corneal structure and hydration in humans and pigs. X-ray scattering data were collected from human and porcine corneas equilibrated with polyethylene glycol (PEG) to varying levels of hydration, to obtain measurements of collagen fibril diameter, interfibrillar spacing (IFS) and intermolecular spacing. Both species showed a strong positive linear correlation between hydration and IFS2 and a nonlinear, bi-phasic relationship between hydration and fibril diameter, whereby fibril diameter increased up to approximately physiological hydration, H = 3.0, with little change thereafter. Above H = 3.0, porcine corneas exhibited a larger fibril diameter than human corneas (p < 0.001). Intermolecular spacing also varied with hydration in a bi-phasic manner but reached a maximum value at a lower hydration (H = 1.5) than fibril diameter. Human corneas displayed a higher intermolecular spacing than porcine corneas at all hydrations (p < 0.0001). Human and porcine corneas required a similar PEG concentration to reach physiological hydration, suggesting that the total fixed charge that gives rise to the swelling pressure is the same. The difference in their structural responses to hydration can be explained by variations in molecular cross-linking and intra/interfibrillar water partitioning. PMID:28592658

A quantitative method to evaluate the donor corneal tissue quality used in a comparative study between two hypothermic preservation media.

PubMed

Parekh, Mohit; Salvalaio, Gianni; Ferrari, Stefano; Amoureux, Marie-Claude; Albrecht, Cecile; Fortier, Denis; Ponzin, Diego

2014-12-01

To standardize a new evaluation technique for calculating the overall quality (OQ) of the donor cornea and validate it using a comparative study of corneas preserved in Optisol-GS and Cornea Cold®. Thirty pairs of donor corneas were selected for a 4 week in vitro comparative study using masked observers. Physiological parameters like thickness, transparency, viable endothelial cell density (VECD) and morphology were transformed to numerical range (0-4) to obtain the OQ. Microbiological examination was performed using Bactec instrument. Students t test showed statistically better results (p < 0.05) from week 3 for thickness, week 2 for transparency and week 1 for morphology and VECD; statistical significance (p < 0.05) was found for OQ from week 2 for the corneas preserved in Cornea Cold® compared to Optisol-GS. Epithelial quality was similar regardless of the medium. Microbiological examination showed absence of aerobic and anaerobic microorganisms in both media. OQ method is efficient, consistent and easy, now validated for comparative studies. Further refinement is necessary for its use at eye-banks, bio-banks and research or transplantation purposes. Cornea Cold® is a promising hypothermic corneal storage medium with preservation time ≤21 days. This permits higher flexibility, evaluation accuracy, longer duration for surgical preparation and ease of transportation.

Regional variation in the refractive-index of the bovine and human cornea.

PubMed

Vasudevan, Balamurali; Simpson, Trefford L; Sivak, Jacob G

2008-10-01

Given the refractive importance of the human cornea, surprisingly little attention has been directed to the study of local variation in corneal refractive-index. This in vitro and in vivo study measures the refractive-index of different portions of the bovine and human cornea. Fifty fresh bovine corneas (obtained from an abattoir) and 10 human subjects were used for the study. The refractive-index of the central, nasal, and temporal corneal epithelium was measured with a bench-top Abbe refractometer in the case of bovine corneas and with a hand-held refractometer with humans. The mean (+/-standard deviation) refractive-indices of the central, nasal, and temporal bovine corneal epithelium were 1.3760 (+/-0.003), 1.3757 (+/-0.002), and 1.3746 (+/-0.002), respectively. Refractive-indices of the anterior and posterior bovine corneal stroma were 1.3731 (+/-0.002) and 1.3708 (+/-0.004), respectively. The mean (+/-standard deviation) refractive-index in the central, nasal, and temporal periphery of the human cornea epithelium were 1.3970 (+/-0.001), 1.3946 (+/-0.001), and 1.3940 (+/-0.001), respectively. There are small local differences in the refractive-index of the bovine and human corneal epithelium and the refractive-index of the epithelium is higher than that of the anterior and posterior stroma of the bovine cornea.

The structural response of the cornea to changes in stromal hydration.

PubMed

Hayes, Sally; White, Tomas; Boote, Craig; Kamma-Lorger, Christina S; Bell, James; Sorenson, Thomas; Terrill, Nick; Shebanova, Olga; Meek, Keith M

2017-06-01

The primary aim of this study was to quantify the relationship between corneal structure and hydration in humans and pigs. X-ray scattering data were collected from human and porcine corneas equilibrated with polyethylene glycol (PEG) to varying levels of hydration, to obtain measurements of collagen fibril diameter, interfibrillar spacing (IFS) and intermolecular spacing. Both species showed a strong positive linear correlation between hydration and IFS 2 and a nonlinear, bi-phasic relationship between hydration and fibril diameter, whereby fibril diameter increased up to approximately physiological hydration, H = 3.0, with little change thereafter. Above H = 3.0, porcine corneas exhibited a larger fibril diameter than human corneas ( p < 0.001). Intermolecular spacing also varied with hydration in a bi-phasic manner but reached a maximum value at a lower hydration ( H = 1.5) than fibril diameter. Human corneas displayed a higher intermolecular spacing than porcine corneas at all hydrations ( p < 0.0001). Human and porcine corneas required a similar PEG concentration to reach physiological hydration, suggesting that the total fixed charge that gives rise to the swelling pressure is the same. The difference in their structural responses to hydration can be explained by variations in molecular cross-linking and intra/interfibrillar water partitioning. © 2017 The Authors.

Expression of pluripotency factors in larval epithelia of the frog Xenopus: Evidence for the presence of cornea epithelial stem cells

PubMed Central

Perry, Kimberly J.; Thomas, Alvin G.; Henry, Jonathan J.

2013-01-01

Understanding the biology of somatic stem cells in self renewing tissues represents an exciting field of study, especially given the potential to harness these cells for tissue regeneration and repair in treating injury and disease. The mammalian cornea contains a population of basal epithelial stem cells involved in cornea homeostasis and repair. Research has been restricted to mammalian systems and little is known about the presence or function of these stem cells in other vertebrates. Therefore, we carried out studies to characterize frog cornea epithelium. Careful examination shows that the Xenopus larval cornea epithelium consists of three distinct layers that include an outer epithelial layer and underlying basal epithelium, in addition to a deeper fibrous layer that contains the main sensory nerve trunks that give rise to numerous branches that extend into these epithelia. These nerves convey sensory and presumably also autonomic innervation to those tissues. The sensory nerves are all derived as branches of the trigeminal nerve/ganglion similar to the situation encountered in mammals, though there appear to be some potentially interesting differences, which are detailed in this paper. We show further that numerous pluripotency genes are expressed by cells in the cornea epithelium, including: sox2, p63, various oct4 homologs, c-myc, klf4 and many others. Antibody localization revealed that p63, a well known mammalian epithelial stem cell marker, was localized strictly to all cells in the basal cornea epithelium. c-myc, was visualized in a smaller subset of basal epithelial cells and adjacent stromal tissue predominately at the periphery of the cornea (limbal zone). Finally, sox2 protein was found to be present throughout all cells of both the outer and basal epithelia, but was much more intensely expressed in a distinct subset of cells that appeared to be either multinucleate or possessed multi-lobed nuclei that are normally located at the periphery of the cornea. Using a thymidine analog (EdU), we were able to label mitotically active cells, which revealed that cell proliferation takes place throughout the cornea epithelium, predominantly in the basal epithelial layer. Species of Xenopus and one other amphibian are unique in their ability to replace a missing lens from cells derived from the basal cornea epithelium. Using EdU we show, as others have previously, that proliferating cells within the cornea epithelium do contribute to the formation of these regenerated lenses. Furthermore, using qPCR we determined that representatives of various pluripotency genes (i.e., sox2, p63 and oct60) are upregulated early during the process of lens regeneration. Antibody labeling showed that the number of sox2 expressing cells increased dramatically within 4 hours following lens removal and these cells were scattered throughout the basal layer of the cornea epithelium. Historically, the process of lens regeneration in Xenopus had been described as one involving transdifferentiation of cornea epithelial cells (i.e., one involving cellular dedifferentiation followed by redifferentiation). Our combined observations provide evidence that a population of stem cells exists within the Xenopus cornea. We hypothesize that the basal epithelium contains oligopotent epithelial stem cells that also represent the source of regenerated lenses in the frog. Future studies will be required to clearly identify the source of these lenses. PMID:23274420

Rabbit cornea microstructure response to changes in intraocular pressure visualized by using nonlinear optical microscopy.

PubMed

Wu, Qiaofeng; Yeh, Alvin T

2008-02-01

To characterize the microstructural response of the rabbit cornea to changes in intraocular pressure (IOP) by using nonlinear optical microscopy (NLOM). Isolated rabbit corneas were mounted on an artificial anterior chamber in series with a manometer and were hydrostatically pressurized by a reservoir. The chamber was mounted on an upright microscope stage of a custom-built NLOM system for corneal imaging without using exogenous stains or dyes. Second harmonic generation in collagen was used to image through the full thickness of the central corneal stroma at IOPs between 5 and 20 mm Hg. Microstructural morphology changes as a function of IOP were used to characterize the depth-dependent response of the central cornea. Regional collagen lamellae architecture through the full thickness of the stroma was specifically imaged as a function of IOP. Hypotensive corneas showed gaps between lamellar structures that decreased in size with increasing IOP. These morphologic features appear to result from interwoven lamellae oriented along the anterior-posterior axis and parallel to the cornea surface. They appear throughout the full thickness and disappear with tension in the anterior but persist in the posterior central cornea, even at hypertensive IOP. NLOM reveals interwoven collagen lamellae sheets through the full thickness of the rabbit central cornea oriented along the anterior-posterior axis and parallel to the surface. The nondestructive nature of NLOM allows 3-dimensional imaging of stromal architecture as a function of IOP in situ. Collagen morphologic features were used as an indirect measure of depth-dependent mechanical response to changes in IOP.

Phototoxic Effect of Topical Fluoroquinolones Administered Before Corneal Crosslinking in a Murine Model.

PubMed

Reviglio, Victor E; Osaba, Matias; Sambuelli, Gabriela; Kuo, Irene C

2017-03-01

Corneal crosslinking by UV light (UV-CXL) has become a popular treatment for keratoconus and corneal ectasia. Fluoroquinolones (FQs), commonly administered topically before UV-CXL, are known to be phototoxic to the skin and lens. The purpose of this study was to investigate phototoxic effects of topical FQ treatment on murine corneas before UV-CXL, in which the corneal epithelium was kept intact. Murine corneas were treated with various antibiotics with or without riboflavin before UV-CXL. At 24 h, the animals were sacrificed, and the corneas were analyzed for histologic evidence of inflammation and apoptosis and for expression of apoptosis markers BAX and caspases 3 and 9 and for expression of matrix metalloproteinase 9 (MMP-9). Spectrofluorometric analysis was performed. Corneas treated with topical FQ with or without riboflavin before UV-CXL showed mild corneal stromal inflammation, apoptosis by both terminal deoxynucleotidyl transferase dUTP nick end labeling staining and increased expression of BAX gene and caspases 3 and 9 by densitometric analysis. Untreated corneas, corneas treated with azithromycin before UV-CXL, and corneas undergoing UV-CXL without any antibiotic or riboflavin pretreatment showed normal histology, no staining for apoptosis, and no increased production of apoptosis markers by polymerase chain reaction. The phototoxic effects of FQs on the cornea may lead surgeons to consider another antibiotic class for prophylaxis against infectious keratitis in UV-CXL. These effects, along with the known cytotoxic effects of FQs independent of UV radiation, may contribute to some of the complications of corneal UV-CXL. Dosage studies may be warranted.

A Simple and Reliable Technique to Orient Donor Corneal Tissue Using the Radial Width of the Surgical Limbus.

PubMed

Aldrich, Benjamin T; Stockman, Adam M; Freiburger, Monica J; Shinkunas, Todd J; Burckart, Kimberlee A; Schmidt, Gregory A; Reed, Cynthia R; Zimmerman, M Bridget; Goins, Kenneth M; Wagoner, Michael D; Greiner, Mark A

2015-12-01

To develop a method based on identification of the widest region of the surgical limbus that can yield quick and accurate orientation of excised human donor corneas. Corneoscleral tissue from donors 49 to 75 years old was marked at the temporal sclera at the time of recovery. Digital images obtained from 20 corneas stored in viewing chambers, retroilluminated and viewed from the endothelial side, were used to quantify the per-degree radial width of the surgical limbus, defined as the distance from the scleral spur to clear cornea. To evaluate differences in radial width among regions, measurements were compared with the intracorneal mean limbal width, and a per-degree z-score was calculated by averaging among corneas. Using images of corneas with the temporal mark masked and the sidedness known, 6 observers were subjected to a blinded trial of 10 corneas to determine the central point of the widest limbal region of each cornea. Compared with the intracorneal mean, the mean radial width of the surgical limbus was greatest in the superior quadrant, and the difference compared with the inferior, nasal, and temporal quadrants was significant (P < 0.0001). The superior region was identified with 100% accuracy in blinded trials. The average absolute difference between the predicted and actual central point of the superior limbus was 9.75 ± 0.30 degrees. The radial width of the surgical limbus is greatest in the superior region of the cornea and can be used as a diagnostic feature to orient donor corneal tissue.

Biological oxygen apparent transmissibility of hydrogel contact lenses with and without organosilicon moieties.

PubMed

Compañ, V; López-Alemany, A; Riande, E; Refojo, M F

2004-01-01

The instrument oxygen transmissibility (IOT) of organosilicon hydrogels, measured by electrochemical procedures, is 5-10 times larger than that of conventional hydrogels. A method is described that allows the estimation of the oxygen tension at the lens-cornea interface for closed- and open-eyelids situations by combining the IOT of the hydrogels and corneal parameters such as corneal thickness, corneal permeability and oxygen flux across the cornea. From these results the biological oxygen apparent transmissibility (BOAT) is obtained, an important parameter which an multiplication with the pressure of oxygen on the external part of the lens gives the oxygen flux onto the cornea. Contact lenses with oxygen transmissibility higher than 100 Dk/t units [1 Dk/t unit=10(-9) [cm(3) O(2) (STp) cm(-2)s(-1)(mmHg)(-1)] posses a large oxygen tension at the lens-cornea interface that substantially reduces the oxygen flux onto the cornea. Lenses whose oxygen transmissibility is lower than 50 Dk/t units allow a rather small oxygen flux onto the cornea under closed eyelids condition that prevent their use for extended wear.

Assessing microstructures of the cornea with Gabor-domain optical coherence microscopy: pathway for corneal physiology and diseases.

PubMed

Tankam, Patrice; He, Zhiguo; Chu, Ying-Ju; Won, Jungeun; Canavesi, Cristina; Lepine, Thierry; Hindman, Holly B; Topham, David J; Gain, Philippe; Thuret, Gilles; Rolland, Jannick P

2015-03-15

Gabor-domain optical coherence microscopy (GD-OCM) was applied ex vivo in the investigation of corneal cells and their surrounding microstructures with particular attention to the corneal endothelium. Experiments using fresh pig eyeballs, excised human corneal buttons from patients with Fuchs' endothelial dystrophy (FED), and healthy donor corneas were conducted. Results show in a large field of view (1  mm×1  mm) high definition images of the different cell types and their surrounding microstructures through the full corneal thickness at both the central and peripheral locations of porcine corneas. Particularly, an image of the endothelial cells lining the bottom of the cornea is highlighted. As compared to healthy human corneas, the corneas of individuals with FED show characteristic microstructural alterations of the Descemet's membrane and increased size and number of keratocytes. The GD-OCM-based imaging system developed may constitute a novel tool for corneal imaging and disease diagnosis. Also, importantly, it may provide insights into the mechanism of corneal physiology and pathology, particularly in diseases of the corneal endothelium.

Comparable change in stromal refractive index of cat and human corneas following blue-IRIS.

PubMed

Wozniak, Kaitlin T; Gearhart, Sara M; Savage, Daniel E; Ellis, Jonathan D; Knox, Wayne H; Huxlin, Krystel R

2017-05-01

Blue intratissue refractive index shaping (blue-IRIS) is a method with potential to correct ocular refraction noninvasively in humans. To date, blue-IRIS has only ever been applied to cat corneas and hydrogels. To test the comparability of refractive index change achievable in cat and human tissues, we used blue-IRIS to write identical phase gratings in ex vivo feline and human corneas. Femtosecond pulses (400 nm) were focused ? 300 ?? ? m below the epithelial surface of excised cat and human corneas and scanned to write phase gratings with lines ? 1 ?? ? m wide, spaced 5 ?? ? m apart, using a scan speed of 5 ?? mm / s . Additional cat corneas were used to test writing at 3 and 7 ?? mm / s in order to document speed dependence of the refractive index change magnitude. The first-order diffraction efficiency was immediately measured and used to calculate the refractive index change attained. Our data show that blue-IRIS induces comparable refractive index changes in feline and human corneas, an essential requirement for further developing its use as a clinical vision correction technique.

Comparable change in stromal refractive index of cat and human corneas following blue-IRIS

NASA Astrophysics Data System (ADS)

Wozniak, Kaitlin T.; Gearhart, Sara M.; Savage, Daniel E.; Ellis, Jonathan D.; Knox, Wayne H.; Huxlin, Krystel R.

2017-05-01

Blue intratissue refractive index shaping (blue-IRIS) is a method with potential to correct ocular refraction noninvasively in humans. To date, blue-IRIS has only ever been applied to cat corneas and hydrogels. To test the comparability of refractive index change achievable in cat and human tissues, we used blue-IRIS to write identical phase gratings in ex vivo feline and human corneas. Femtosecond pulses (400 nm) were focused ˜300 μm below the epithelial surface of excised cat and human corneas and scanned to write phase gratings with lines ˜1 μm wide, spaced 5 μm apart, using a scan speed of 5 mm/s. Additional cat corneas were used to test writing at 3 and 7 mm/s in order to document speed dependence of the refractive index change magnitude. The first-order diffraction efficiency was immediately measured and used to calculate the refractive index change attained. Our data show that blue-IRIS induces comparable refractive index changes in feline and human corneas, an essential requirement for further developing its use as a clinical vision correction technique.

In vivo 3D measurement of moxifloxacin and gatifloxacin distributions in the mouse cornea using multiphoton microscopy

NASA Astrophysics Data System (ADS)

Lee, Seunghun; Lee, Jun Ho; Park, Jin Hyoung; Yoon, Yeoreum; Chung, Wan Kyun; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

2016-05-01

Moxifloxacin and gatifloxacin are fourth-generation fluoroquinolone antibiotics used in the clinic to prevent or treat ocular infections. Their pharmacokinetics in the cornea is usually measured from extracted ocular fluids or tissues, and in vivo direct measurement is difficult. In this study multiphoton microscopy (MPM), which is a 3D optical microscopic technique based on multiphoton fluorescence, was applied to the measurement of moxifloxacin and gatifloxacin distribution in the cornea. Intrinsic multiphoton fluorescence properties of moxifloxacin and gatifloxacin were characterized, and their distributions in mouse cornea in vivo were measured by 3D MPM imaging. Both moxifloxacin and gatifloxacin had similar multiphoton spectra, while moxifloxacin had stronger fluorescence than gatifloxacin. MPM imaging of mouse cornea in vivo showed (1) moxifloxacin had good penetration through the superficial corneal epithelium, while gatifloxacin had relatively poor penetration, (2) both ophthalmic solutions had high intracellular distribution. In vivo MPM results were consistent with previous studies. This study demonstrates the feasibility of MPM as a method for in vivo direct measurement of moxifloxacin and gatifloxacin in the cornea.

Outcomes of shunt tube coverage with glycerol preserved cornea versus pericardium.

PubMed

Wigton, Eric; C Swanner, Jason; Joiner, Wade; Feldman, Alex; McGwin, Gerald; Huisingh, Carrie; Curcio, Christine A; Girkin, Christopher A

2014-01-01

Pericardium is a biomaterial widely used for covering the outflow tubes of glaucoma drainage devices. Recently, glycerol preserved cornea has been introduced as an alternative that offers durability and improved cosmesis because of its clarity. We retrospectively reviewed 262 patients in the University of Alabama Birmingham Glaucoma Service who underwent shunt procedures using either cornea tissue or pericardium to cover the tube. The primary outcome measure was the number of erosions of the covering material. Nine out of 101 (8.9%) patients in the pericardium covered group experienced an erosion compared with 3 out of 161 (1.9%) in the cornea covered group. A significant difference was reached with P=0.0125. Median follow-up was 440 days for the cornea group and 331 days for the pericardium group. The type of glaucoma (primary open-angle glaucoma vs. secondary glaucoma) was not associated with the risk of erosion (odds ratio, 0.501; 95% confidence interval, 0.204-1.234). The median time to exposure was 252 days in the pericardium group and 440 days in the cornea group (P=0.0017).

In vivo 3D measurement of moxifloxacin and gatifloxacin distributions in the mouse cornea using multiphoton microscopy

PubMed Central

Lee, Seunghun; Lee, Jun Ho; Park, Jin Hyoung; Yoon, Yeoreum; Chung, Wan Kyun; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

2016-01-01

Moxifloxacin and gatifloxacin are fourth-generation fluoroquinolone antibiotics used in the clinic to prevent or treat ocular infections. Their pharmacokinetics in the cornea is usually measured from extracted ocular fluids or tissues, and in vivo direct measurement is difficult. In this study multiphoton microscopy (MPM), which is a 3D optical microscopic technique based on multiphoton fluorescence, was applied to the measurement of moxifloxacin and gatifloxacin distribution in the cornea. Intrinsic multiphoton fluorescence properties of moxifloxacin and gatifloxacin were characterized, and their distributions in mouse cornea in vivo were measured by 3D MPM imaging. Both moxifloxacin and gatifloxacin had similar multiphoton spectra, while moxifloxacin had stronger fluorescence than gatifloxacin. MPM imaging of mouse cornea in vivo showed (1) moxifloxacin had good penetration through the superficial corneal epithelium, while gatifloxacin had relatively poor penetration, (2) both ophthalmic solutions had high intracellular distribution. In vivo MPM results were consistent with previous studies. This study demonstrates the feasibility of MPM as a method for in vivo direct measurement of moxifloxacin and gatifloxacin in the cornea. PMID:27138688

Material Properties from Air Puff Corneal Deformation by Numerical Simulations on Model Corneas

PubMed Central

Dorronsoro, Carlos; de la Hoz, Andrés; Marcos, Susana

2016-01-01

Objective To validate a new method for reconstructing corneal biomechanical properties from air puff corneal deformation images using hydrogel polymer model corneas and porcine corneas. Methods Air puff deformation imaging was performed on model eyes with artificial corneas made out of three different hydrogel materials with three different thicknesses and on porcine eyes, at constant intraocular pressure of 15 mmHg. The cornea air puff deformation was modeled using finite elements, and hyperelastic material parameters were determined through inverse modeling, minimizing the difference between the simulated and the measured central deformation amplitude and central-peripheral deformation ratio parameters. Uniaxial tensile tests were performed on the model cornea materials as well as on corneal strips, and the results were compared to stress-strain simulations assuming the reconstructed material parameters. Results The measured and simulated spatial and temporal profiles of the air puff deformation tests were in good agreement (< 7% average discrepancy). The simulated stress-strain curves of the studied hydrogel corneal materials fitted well the experimental stress-strain curves from uniaxial extensiometry, particularly in the 0–0.4 range. Equivalent Young´s moduli of the reconstructed material properties from air-puff were 0.31, 0.58 and 0.48 MPa for the three polymer materials respectively which differed < 1% from those obtained from extensiometry. The simulations of the same material but different thickness resulted in similar reconstructed material properties. The air-puff reconstructed average equivalent Young´s modulus of the porcine corneas was 1.3 MPa, within 18% of that obtained from extensiometry. Conclusions Air puff corneal deformation imaging with inverse finite element modeling can retrieve material properties of model hydrogel polymer corneas and real corneas, which are in good correspondence with those obtained from uniaxial extensiometry, suggesting that this is a promising technique to retrieve quantitative corneal biomechanical properties. PMID:27792759

The three-dimensional microanatomy of the rabbit and human cornea. A chemical and mechanical microdissection-SEM approach

PubMed Central

OJEDA, JOSÉ L.; VENTOSA, JUAN A.; PIEDRA, SONSOLES

2001-01-01

The three-dimensional (3D) microanatomy of the cornea is the major determinant of its optical and mechanical properties. Scanning electron microscopy (SEM) is the most commonly used method to obtain information on the overall 3D microanatomy of organs. However, SEM has not been successful in revealing the 3D microanatomy of the cornea, because the interior of the cornea is too compact to be explored by the electron beam. In this study, the 3D organisation of the cells and extracellular materials of human and rabbit corneas was examined after exposure by HCl and NaOH digestion, and by microdissection by the adhesive tape method. In the cornea of both species, all epithelial cells exhibited microplicae regardless of their location. This raises doubts about the tear film-holding role assigned to the microplicae of the superficial cells. Human and rabbit corneas differed in the collagen fibre patterns of the epithelial basement membranes. The 3D organisation of the stromal lamellae was similar in both species. In humans and rabbits, the keratocytes showed similar 3D features. However, the surface of human keratocytes located near Descemet's membrane exhibited small fenestrations that were not present in the rabbit keratocytes. The pattern of keratocyte innervation by the stromal neural plexus and 3D keratocyte microanatomy confirms that keratocytes form a large intercommunicating network within the corneal stroma. Two morphologically discrete subpopulations of keratocytes located at different stromal levels were identified in both human and rabbit corneas, suggesting that keratocytes are not functionally homogeneous. In addition, the density of the stromal neural plexus appeared to be greater in rabbits than in humans. Clear differences between human and rabbit corneas were observed in the collagen arrangement in Descemet's membrane, which may reflect their different biomechanical requirements. PMID:11760887

Ex vivo rabbit and human corneas as models for bacterial and fungal keratitis.

PubMed

Pinnock, Abigail; Shivshetty, Nagaveni; Roy, Sanhita; Rimmer, Stephen; Douglas, Ian; MacNeil, Sheila; Garg, Prashant

2017-02-01

In the study of microbial keratitis, in vivo animal models often require a large number of animals, and in vitro monolayer cell culture does not maintain the three-dimensional structure of the tissues or cell-to-cell communication of in vivo models. Here, we propose reproducible ex vivo models of single- and dual-infection keratitis as an alternative to in vivo and in vitro models. Excised rabbit and human corneoscleral rims maintained in organ culture were infected using 10 8 cells of Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans or Fusarium solani. The infection was introduced by wounding with a scalpel and exposing corneas to the microbial suspension or by intrastromal injection. Post-inoculation, corneas were maintained for 24 and 48 h at 37 °C. After incubation, corneas were either homogenised to determine colony-forming units (CFU)/cornea or processed for histological examination using routine staining methods. Single- and mixed-species infections were compared. We observed a significant increase in CFU after 48 h compared to 24 h with S. aureus and P. aeruginosa. However, no such increase was observed in corneas infected with C. albicans or F. solani. The injection method yielded an approximately two- to 100-fold increase (p < 0.05) in the majority of organisms from infected corneas. Histology of the scalpel-wounded and injection models indicated extensive infiltration of P. aeruginosa throughout the entire cornea, with less infiltration observed for S. aureus, C. albicans and F. solani. The models also supported dual infections. Both scalpel wounding and injection methods are suitable for inducing infection of ex vivo rabbit and human cornea models. These simple and reproducible models will be useful as an alternative to in vitro and in vivo models for investigating the detection and treatment of microbial keratitis, particularly when this might be due to two infective organisms.

Diclofenac sodium delivery to the eye: in vitro evaluation of novel solid lipid nanoparticle formulation using human cornea construct.

PubMed

Attama, Anthony A; Reichl, Stephan; Müller-Goymann, Christel C

2008-05-01

Solid lipid nanoparticles (SLNs) were prepared with a combination of homolipid from goat (goat fat) and phospholipid, and evaluated for diclofenac sodium (DNa) delivery to the eye using bio-engineered human cornea, produced from immortalized human corneal endothelial cells (HENC), stromal fibroblasts and epithelial cells CEPI 17 CL 4. Encapsulation efficiency was high and sustained release of DNa and high permeation through the bio-engineered cornea were achieved. Results obtained in this work showed that permeation of DNa through the cornea construct was improved by formulation as SLN modified with phospholipid.

Factors influencing the virological testing of cornea donors.

PubMed

Röck, Tobias; Beck, Robert; Jürgens, Stefan; Bartz-Schmidt, Karl Ulrich; Bramkamp, Matthias; Thaler, Sebastian; Röck, Daniel

2017-11-01

To assess the influence of donor, environment, and logistical factors on the results of virological testing of blood samples from cornea donors.Data from 670 consecutive cornea donors were analyzed retrospectively. Logistic regression analysis was used to assess the influence of different factors on the results of virological testing of blood samples from cornea donors.The mean annual rate of donors with serology-reactive or not evaluable result was 14.8% (99 of 670) (range 11.9%-16.9%). The cause of donor death by cancer increased the risk of serology-reactive or not evaluable result (P = .0300). Prolonged time between death and post mortem blood removal was associated with a higher rate of serology-reactive or not evaluable result (P < .0001). Mean monthly temperature including warmer months, differentiating between septic and aseptic donors, sex, and donor age had no significant impact on the results of virological testing of blood samples from cornea donors.The cause of donor death by cancer and a prolonged time between death and post mortem blood removal seem to be mainly responsible for serology-reactive or not evaluable result of blood samples from cornea donors. The percentage of discarded corneas caused by serology-reactive or not evaluable result may be reduced by shortening the period of time between death and post mortem blood removal. Copyright © 2017 The Authors. Published by Wolters Kluwer Health, Inc. All rights reserved.

On the barrier properties of the cornea: a microscopy study of the penetration of fluorescently labeled nanoparticles, polymers, and sodium fluorescein.

PubMed

Mun, Ellina A; Morrison, Peter W J; Williams, Adrian C; Khutoryanskiy, Vitaliy V

2014-10-06

Overcoming the natural defensive barrier functions of the eye remains one of the greatest challenges of ocular drug delivery. Cornea is a chemical and mechanical barrier preventing the passage of any foreign bodies including drugs into the eye, but the factors limiting penetration of permeants and nanoparticulate drug delivery systems through the cornea are still not fully understood. In this study, we investigate these barrier properties of the cornea using thiolated and PEGylated (750 and 5000 Da) nanoparticles, sodium fluorescein, and two linear polymers (dextran and polyethylene glycol). Experiments used intact bovine cornea in addition to bovine cornea de-epithelialized or tissues pretreated with cyclodextrin. It was shown that corneal epithelium is the major barrier for permeation; pretreatment of the cornea with β-cyclodextrin provides higher permeation of low molecular weight compounds, such as sodium fluorescein, but does not enhance penetration of nanoparticles and larger molecules. Studying penetration of thiolated and PEGylated (750 and 5000 Da) nanoparticles into the de-epithelialized ocular tissue revealed that interactions between corneal surface and thiol groups of nanoparticles were more significant determinants of penetration than particle size (for the sizes used here). PEGylation with polyethylene glycol of a higher molecular weight (5000 Da) allows penetration of nanoparticles into the stroma, which proceeds gradually, after an initial 1 h lag phase.

Wound-Healing Studies in Cornea and Skin: Parallels, Differences and Opportunities

PubMed Central

Bukowiecki, Anne; Hos, Deniz; Cursiefen, Claus; Eming, Sabine A.

2017-01-01

The cornea and the skin are both organs that provide the outer barrier of the body. Both tissues have developed intrinsic mechanisms that protect the organism from a wide range of external threats, but at the same time also enable rapid restoration of tissue integrity and organ-specific function. The easy accessibility makes the skin an attractive model system to study tissue damage and repair. Findings from skin research have contributed to unravelling novel fundamental principles in regenerative biology and the repair of other epithelial-mesenchymal tissues, such as the cornea. Following barrier disruption, the influx of inflammatory cells, myofibroblast differentiation, extracellular matrix synthesis and scar formation present parallel repair mechanisms in cornea and skin wound healing. Yet, capillary sprouting, while pivotal in proper skin wound healing, is a process that is rather associated with pathological repair of the cornea. Understanding the parallels and differences of the cellular and molecular networks that coordinate the wound healing response in skin and cornea are likely of mutual importance for both organs with regard to the development of regenerative therapies and understanding of the disease pathologies that affect epithelial-mesenchymal interactions. Here, we review the principal events in corneal wound healing and the mechanisms to restore corneal transparency and barrier function. We also refer to skin repair mechanisms and their potential implications for regenerative processes in the cornea. PMID:28604651

Corneal delivery of besifloxacin using rapidly dissolving polymeric microneedles.

PubMed

Bhatnagar, Shubhmita; Saju, Amala; Cheerla, Krishna Deepthi; Gade, Sudeep Kumar; Garg, Prashant; Venuganti, Venkata Vamsi Krishna

2018-06-01

Penetration of antibiotics into and through the cornea is a major limiting factor in the treatment of ocular infections. Several strategies are in vogue to overcome this limitation such as use of fortified drops, gels, and subconjunctival injections. Here, we present the fabrication of rapidly dissolving polymeric microneedle array to effectively deliver besifloxacin through the cornea. Microneedles were prepared using polyvinyl alcohol and polyvinyl pyrrolidone by the micromolding technique. The model fluoroquinolone antibiotic, besifloxacin, was loaded in 36 microneedles arranged in a 6 × 6 array format within a 1 cm 2 area. The average height and base width of microneedles was 961 ± 27 and 366 ± 16 μm, respectively. Each microneedle array contained 103.4 ± 8.5 μg of besifloxacin. Cryosectioning and confocal microscopy of excised human cornea revealed that microneedles penetrated to a depth of up to 200 μm. Microneedles were found to completely dissolve in the cornea within 5 min. Application of microneedles for 5 min significantly (p < 0.05) improved the besifloxacin deposition and permeation through the cornea compared with free besifloxacin solution. Similarly, besifloxacin-loaded microneedles showed greater antibacterial activity in Staphylococcus aureus-infected cornea in comparison to free besifloxacin solution. Taken together, rapidly dissolving microneedles can be developed to effectively deliver besifloxacin to treat bacterial infections in the cornea and eye.

Spherical aberrations of human astigmatic corneas.

PubMed

Zhao, Huawei; Dai, Guang-Ming; Chen, Li; Weeber, Henk A; Piers, Patricia A

2011-11-01

To evaluate whether the average spherical aberration of human astigmatic corneas is statistically equivalent to human nonastigmatic corneas. Spherical aberrations of 445 astigmatic corneas prior to laser vision correction were retrospectively investigated to determine Zernike coefficients for central corneal areas 6 mm in diameter using CTView (Sarver and Associates). Data were divided into groups according to cylinder power (0.01 to 0.25 diopters [D], 0.26 to 0.75 D, 0.76 to 1.06 D, 1.07 to 1.53 D, 1.54 to 2.00 D, and >2.00 D) and according to age by decade. Spherical aberrations were correlated with age and astigmatic power among groups and the entire population. Statistical analyses were conducted, and P<.05 was considered statistically significant. Mean patient age was 42.6±11 years. Astigmatic corneas had an average astigmatic power of 0.78±0.58 D and mean spherical aberration was 0.25±0.13 μm for the entire population and approximately the same (0.27 μm) for individual groups, ranging from 0.23 to 0.29 μm (P>.05 for all tested groups). Mean spherical aberration of astigmatic corneas was not correlated significantly with cylinder power or age (P>.05). Spherical aberrations are similar to those of nonastigmatic corneas, permitting the use of these additional data in the design of aspheric toric intra-ocular lenses. Copyright 2011, SLACK Incorporated.

Factors influencing the virological testing of cornea donors

PubMed Central

Röck, Tobias; Beck, Robert; Jürgens, Stefan; Bartz-Schmidt, Karl Ulrich; Bramkamp, Matthias; Thaler, Sebastian; Röck, Daniel

2017-01-01

Abstract To assess the influence of donor, environment, and logistical factors on the results of virological testing of blood samples from cornea donors. Data from 670 consecutive cornea donors were analyzed retrospectively. Logistic regression analysis was used to assess the influence of different factors on the results of virological testing of blood samples from cornea donors. The mean annual rate of donors with serology-reactive or not evaluable result was 14.8% (99 of 670) (range 11.9%–16.9%). The cause of donor death by cancer increased the risk of serology-reactive or not evaluable result (P = .0300). Prolonged time between death and post mortem blood removal was associated with a higher rate of serology-reactive or not evaluable result (P < .0001). Mean monthly temperature including warmer months, differentiating between septic and aseptic donors, sex, and donor age had no significant impact on the results of virological testing of blood samples from cornea donors. The cause of donor death by cancer and a prolonged time between death and post mortem blood removal seem to be mainly responsible for serology-reactive or not evaluable result of blood samples from cornea donors. The percentage of discarded corneas caused by serology-reactive or not evaluable result may be reduced by shortening the period of time between death and post mortem blood removal. PMID:29381929

Evaluation of active and passive transport processes in corneas extracted from preserved rabbit eyes.

PubMed

Majumdar, Soumyajit; Hingorani, Tushar; Srirangam, Ramesh

2010-04-01

In vitro transcorneal permeability studies are an important screening tool in drug development. The objective of this research is to examine the feasibility of using corneas isolated from preserved rabbit eyes as a model for permeability evaluation. Eyes from male New Zealand White rabbits were used immediately or were stored overnight in phosphate-buffered saline (PBS) or Hanks balanced salt solution (HBSS) over wet ice. Integrity of isolated corneas was evaluated by measuring the TEER and by determining the permeability of paracellular and transcellular markers. Active transport was assessed by measuring transcorneal permeability of selected amino acids. Esterase activity was estimated using p-nitrophenyl assay. In all cases, corneas from freshly enucleated eyes were compared to those isolated from the day-old preserved eyes. Transcellular and paracellular passive diffusion was not affected by the storage medium and observed to be similar in the fresh and preserved eye models. However, amino acid transporters demonstrated lower functional activity in corneas excised from eyes preserved in PBS. Moreover, preserved eyes displayed almost 1.5-fold lower esterase activity in the corneal tissue. Thus, corneas isolated from day-old eyes, preserved in HBSS, closely mimics freshly excised rabbit corneas in terms of both active and passive transport characteristics but possesses slightly reduced enzymatic activity. 2009 Wiley-Liss, Inc. and the American Pharmacists Association

Wound-Healing Studies in Cornea and Skin: Parallels, Differences and Opportunities.

PubMed

Bukowiecki, Anne; Hos, Deniz; Cursiefen, Claus; Eming, Sabine A

2017-06-12

The cornea and the skin are both organs that provide the outer barrier of the body. Both tissues have developed intrinsic mechanisms that protect the organism from a wide range of external threats, but at the same time also enable rapid restoration of tissue integrity and organ-specific function. The easy accessibility makes the skin an attractive model system to study tissue damage and repair. Findings from skin research have contributed to unravelling novel fundamental principles in regenerative biology and the repair of other epithelial-mesenchymal tissues, such as the cornea. Following barrier disruption, the influx of inflammatory cells, myofibroblast differentiation, extracellular matrix synthesis and scar formation present parallel repair mechanisms in cornea and skin wound healing. Yet, capillary sprouting, while pivotal in proper skin wound healing, is a process that is rather associated with pathological repair of the cornea. Understanding the parallels and differences of the cellular and molecular networks that coordinate the wound healing response in skin and cornea are likely of mutual importance for both organs with regard to the development of regenerative therapies and understanding of the disease pathologies that affect epithelial-mesenchymal interactions. Here, we review the principal events in corneal wound healing and the mechanisms to restore corneal transparency and barrier function. We also refer to skin repair mechanisms and their potential implications for regenerative processes in the cornea.

Effect of corneal inhomogeneity on the mechanical behavior of the eye

NASA Astrophysics Data System (ADS)

Stein, A. A.; Moiseeva, I. N.

2018-05-01

The effect of spatial inhomogeneity of the effective cornea stiffness distribution on the mechanical properties of the eye is investigated on the basis of the two-component model of the eyeball, in which the cornea is represented by a momentless deformable, linearly elastic surface and the scleral region by an elastic element that responds to changes in intraocular pressure by changes in volume. The approach used makes it possible to consider within the same model both the natural corneal inhomogeneity and mechanical consequences of local cornea weakening owing to surgical procedures. The dependences on changes in intraocular pressure of parameters that characterize deformation properties of both the cornea (apex displacement) and the eyeball as a whole (change in intraocular volume) are obtained. For moderate inhomogeneity they differ from the same dependences for the homogenous cornea with effective stiffness equal to the average value for the corresponding inhomogeneous distribution only slightly. However, if the effective stiffness amplitude is very high, corneal inhomogeneity discernibly affects the integral response of the cornea and the eyeball as a whole to changes in pressure. The effect of inhomogeneity on the data of tonometry also mainly depends on the average effective corneal stiffness. The difference between the tonometric and true pressures increases with surgical cornea weakening in the apical region for both Schiøtz and Maklakoff tonometers.

Synthetic neurotensin analogues are nontoxic analgesics for the rabbit cornea.

PubMed

Kim, Charles; Barbut, Denise; Heinemann, Murk H; Pasternak, Gavril; Rosenblatt, Mark I

2014-05-13

To characterize the analgesic potency and toxicity of topical synthetic neurotensin analogues, and localize neurotensin receptors in the cornea and trigeminal ganglion. Cochet-Bonnet esthesiometry was performed on the rabbit cornea to test the analgesic dose response and duration of effect for two synthetic neurotensin analogues: NT71 and NT72. Receptors for neurotensin were localized in the murine cornea and trigeminal ganglion using quantitative PCR (qPCR), Western blotting, and immunohistochemistry. In vitro toxicity of NT71, NT72, and sodium channel blockers was evaluated using cytotoxicity, single-cell migration, and scratch closure assays performed on rabbit corneal epithelial cells. In vivo toxicity of these agents was assessed using a rabbit laser phototherapeutic keratectomy (PTK) model and histology. NT71 and NT72 induced potent analgesic effects on the rabbit cornea at concentrations between 1.0 and 2.5 mg/mL, lasting up to 180 minutes. A site-specific distribution of neurotensin receptors was observed in the murine cornea and trigeminal ganglion. NT71 and NT72 did not cause any significant in vitro or in vivo toxicity, in contrast to sodium channel blockers. Synthetic neurotensin analogues are potent analgesics that avoid the toxicities associated with established topical analgesic agents. Receptors for neurotensin are present in both the cornea and trigeminal ganglion. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

Delivery of Riboflavin-5'-Monophosphate Into the Cornea: Can Liposomes Provide Any Enhancement Effects?

PubMed

Kandzija, Neva; Khutoryanskiy, Vitaliy V

2017-10-01

Keratoconus is a progressive condition caused by the thinning of the cornea, which eventually deforms the front surface of the eye into a cone shape leading to ghosting, multiple images, glare, and several other vision problems. Currently, keratoconus is treated with UV-induced riboflavin (Rb)-mediated collagen cross-linking, which requires a physical removal of the corneal epithelium under topical anesthesia. This study reports the penetration of Rb and its more water-soluble form, riboflavin-5'-monophosphate (RbP), into the bovine cornea ex vivo. Using ex vivo bovine corneal tissues and 0.8 mg/mL drug solutions in phosphate buffer, it was established that RbP penetration into the cornea within 3 h of diffusion experiment was greater (17.3 ± 0.8 μg) compared with Rb (10.4 ± 4.2 μg). In the cornea, RbP was found to convert to Rb, which is mediated with enzymes present in this tissue. Several formulations including the conventional and propylene glycol-containing liposomes with encapsulated RbP have been developed, and their effect on the drug penetration into the bovine cornea was evaluated. Encapsulation of RbP into the liposomes did not provide any statistically significant improvement in the penetration of RbP into the cornea. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Changes in shape and astigmatism of total, anterior, and posterior cornea after long versus short clear corneal incision cataract surgery.

PubMed

Hayashi, Ken; Yoshida, Motoaki; Hirata, Akira; Yoshimura, Koichi

2018-01-01

To compare changes in the shape and astigmatism of the total, anterior, and posterior cornea between eyes with long-length clear corneal incisions (CCIs) and eyes with short-length CCIs in cataract surgery. Hayashi Eye Hospital, Fukuoka, Japan. Prospective case series. Both eyes of patients having phacoemulsification with temporal CCIs were randomized to have a long-length (≥1.75 mm) or short-length (<1.75 mm) CCI. Corneal astigmatic changes were decomposed to vertical/horizontal (Jackson cross-cylinder, axes at 180 degrees and 90 degrees [J0]) and oblique changes (Jackson cross-cylinder, axes at 45 degrees and 135 degrees [J45]) using power vector analysis. Corneal shape changes were assessed using the average of the difference map on videokeratography 2 days and 2, 4, and 8 weeks postoperatively. The mean J0 and J45 values of the total cornea in the 120 eyes evaluated were significantly greater in the long CCI group than in the short CCI group at all follow-ups (P ≤ .0290). The videokeratography showed a wedge-shaped flattening in the total and anterior cornea and a steepening in the posterior cornea around the CCI 2 days postoperatively in both groups. This wound-related flattening of the total and anterior cornea rapidly reduced but persisted until 8 weeks, whereas the steepening of the posterior cornea disappeared within 4 weeks. These changes extended closer to the central cornea in the long CCI group than in the short CCI group. Corneal astigmatic changes were significantly greater after long CCI than after short CCI. The wound-related shape changes occurred immediately postoperatively but rapidly diminished. Copyright © 2018 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

Establishing Corneal Cross-Linking With Riboflavin and UV-A in the Mouse Cornea In Vivo: Biomechanical Analysis.

PubMed

Hammer, Arthur; Kling, Sabine; Boldi, Marc-Olivier; Richoz, Olivier; Tabibian, David; Randleman, J Bradley; Hafezi, Farhad

2015-10-01

To establish corneal cross-linking (CXL) with riboflavin and UV-A in in the mouse cornea in vivo and to develop tools to measure the biomechanical changes observed. A total of 55 male C57BL/6 wild-type mice (aged 5 weeks) were divided into 14 groups. Standard CXL parameters were adapted to the anatomy of the mouse cornea, and riboflavin concentration (0.1%-0.5%) and fluence series (0.09-5.4 J/cm²) were performed on the assumption of the endothelial damage thresholds. Untreated and riboflavin only corneas were used as controls. Animals were killed at 30 minutes and at 1 month after CXL. Corneas were harvested. Two-dimensional (2D) biomechanical testing was performed using a customized corneal holder in a commercially available stress-strain extensometer/indenter. Both elastic and viscoelastic analyses were performed. Statistical inference was performed using t-tests and specific mathematical models fitted to the experimental stress-strain and stress-relaxation data. Adjusted P values by the method of Benjamini and Hochberg are reported. For all CXL treatment groups, stress-relaxation showed significant differences (P < 0.0001) after 120 seconds of constant strain application, with cross-linked corneas maintaining a higher stress (441 ± 40 kPa) when compared with controls (337 ± 39 kPa). Stress-strain analysis confirmed these findings but was less sensitive to CXL-induced changes: at 0.5% of strain, cross-linked corneas remained at higher stress (778 ± 111 kPa) when compared with controls (659 ± 121 kPa). Cross-linking was induced in the mouse cornea in vivo, and its biomechanical effect successfully measured. This could create opportunities to study molecular pathways of CXL in transgenic mice.

Effect of femtosecond and microkeratome flaps creation on the cornea biomechanics during laser in situ keratomileusis: one year follow-up

PubMed Central

Sun, Qian; Deng, Zheng-Zheng; Zhou, Yue-Hua; Zhang, Jing; Peng, Xiao-Yan

2016-01-01

AIM To compare the corneal biomechanical outcomes at one year after laser in situ keratomileusis (LASIK) with the flaps created by Ziemer and Moria M2 microkeratome with 110 head and -20 blade. METHODS Totally 100 eyes of 50 consecutive patients were enrolled in this prospective study and divided into two groups for corneal flaps created by ZiemerFemto LDV and Moria M2 microkeratome with 110 head and -20 blade. Corneal biomechanical properties including cornea resistance factor (CRF) and cornea hysteresis (CH) were measured before and 1, 3, 6, 12mo after surgery by ocular response analyzer. Central cornea thickness and corneal flap thickness were measured by optical coherence tomography. RESULTS The ablation depth (P=0.693), residual corneal thickness (P=0.453), and postoperative corneal curvature (P=0.264) were not significant different between Ziemer group and Moria 110-20 group after surgery. The residual stromal bed thickness, corneal flap thickness, CH and CRF at 12mo after surgery were significant different between Ziemer group and Moria 110-20 group (P<0.01);Ziemer group gained better corneal biomechanical results. The CRF and CH increased gradually from 1 to 12mo after surgery in Ziemer group, increased from 1 to 6mo but decreased from 6 to 12mo in Moria 110-20 group. Both CRF and CH at one year after surgery increased with the increasing of residual cornea thickness; pre-LASIK CRF, CRF also increased with residual stromal bed thickness, while CH decreased with the increasing of pre-LASIK intraocular pressure and cornea flap thickness (P<0.01). CONCLUSION In one year follow-up, femtosecond laser can provide better cornea flaps with stable cornea biomechanics than mechanical microkeratome. PMID:27803856

Effect of trapping vascular endothelial growth factor-A in a murine model of dry eye with inflammatory neovascularization.

PubMed

Kwon, Jin Woo; Choi, Jin A; Shin, Eun Young; La, Tae Yoon; Jee, Dong Hyun; Chung, Yeon Woong; Cho, Yang Kyung

2016-01-01

To evaluate whether trapping vascular endothelial growth factor A (VEGF-A) would suppress angiogenesis and inflammation in dry eye corneas in a murine corneal suture model. We established two groups of animals, one with non-dry eyes and the other with induced dry eyes. In both groups, a corneal suture model was used to induce inflammation and neovascularization. Each of two groups was again divided into three subgroups according to the treatment; subgroup I (aflibercept), subgroup II (dexamethasone) and subgroup III (phosphate buffered saline, PBS). Corneas were harvested and immunohistochemical staining was performed to compare the extents of neovascularization and CD11b+ cell infiltration. Real-time polymerase chain reaction was performed to quantify the expression of inflammatory cytokines and VEGF-A in the corneas. Trapping VEGF-A with aflibercept resulted in significantly decreased angiogenesis and inflammation compared with the dexamethasone and PBS treatments in the dry eye corneas (all P <0.05), but with no such effects in non-dry eyes. The anti-inflammatory and anti-angiogenic effects of VEGF-A trapping were stronger than those of dexamethasone in both dry eye and non-dry eye corneas (all P <0.05). The levels of RNA expression of VEGF-A, TNF-alpha, and IL-6 in the aflibercept subgroup were significantly decreased compared with those in the PBS subgroup in the dry eye group. Compared with non-dry eye corneas, dry eye corneas have greater amounts of inflammation and neovascularization and also have a more robust response to anti-inflammatory and anti-angiogenic agents after ocular surface surgery. Trapping VEGF-A is effective in decreasing both angiogenesis and inflammation in dry eye corneas after ocular surface surgery.

The Structural Role of Elastic Fibers in the Cornea Investigated Using a Mouse Model for Marfan Syndrome

PubMed Central

White, Tomas L.; Lewis, Philip; Hayes, Sally; Fergusson, James; Bell, James; Farinha, Luis; White, Nick S.; Pereira, Lygia V.; Meek, Keith M.

2017-01-01

Purpose The presence of fibrillin-rich elastic fibers in the cornea has been overlooked in recent years. The aim of the current study was to elucidate their functional role using a mouse model for Marfan syndrome, defective in fibrillin-1, the major structural component of the microfibril bundles that constitute most of the elastic fibers. Methods Mouse corneas were obtained from animals with a heterozygous fibrillin-1 mutation (Fbn1+/−) and compared to wild type controls. Corneal thickness and radius of curvature were calculated using optical coherence tomography microscopy. Elastic microfibril bundles were quantified and visualized in three-dimensions using serial block face scanning electron microscopy. Transmission electron microscopy was used to analyze stromal ultrastructure and proteoglycan distribution. Center-to-center average interfibrillar spacing was determined using x-ray scattering. Results Fbn1+/− corneas were significantly thinner than wild types and displayed a higher radius of curvature. In the Fbn1+/− corneas, elastic microfibril bundles were significantly reduced in density and disorganized compared to wild-type controls, in addition to containing a higher average center-to-center collagen interfibrillar spacing in the center of the cornea. No other differences were detected in stromal ultrastructure or proteoglycan distribution between the two groups. Proteoglycan side chains appeared to colocalize with the microfibril bundles. Conclusions Elastic fibers have an important, multifunctional role in the cornea as highlighted by the differences observed between Fbn1+/− and wild type animals. We contend that the presence of normal quantities of structurally organized elastic fibers are required to maintain the correct geometry of the cornea, which is disrupted in Marfan syndrome. PMID:28395026

Effect of trapping vascular endothelial growth factor-A in a murine model of dry eye with inflammatory neovascularization

PubMed Central

Kwon, Jin Woo; Choi, Jin A; Shin, Eun Young; La, Tae Yoon; Jee, Dong Hyun; Chung, Yeon Woong; Cho, Yang Kyung

2016-01-01

AIM To evaluate whether trapping vascular endothelial growth factor A (VEGF-A) would suppress angiogenesis and inflammation in dry eye corneas in a murine corneal suture model. METHODS We established two groups of animals, one with non-dry eyes and the other with induced dry eyes. In both groups, a corneal suture model was used to induce inflammation and neovascularization. Each of two groups was again divided into three subgroups according to the treatment; subgroup I (aflibercept), subgroup II (dexamethasone) and subgroup III (phosphate buffered saline, PBS). Corneas were harvested and immunohistochemical staining was performed to compare the extents of neovascularization and CD11b+ cell infiltration. Real-time polymerase chain reaction was performed to quantify the expression of inflammatory cytokines and VEGF-A in the corneas. RESULTS Trapping VEGF-A with aflibercept resulted in significantly decreased angiogenesis and inflammation compared with the dexamethasone and PBS treatments in the dry eye corneas (all P<0.05), but with no such effects in non-dry eyes. The anti-inflammatory and anti-angiogenic effects of VEGF-A trapping were stronger than those of dexamethasone in both dry eye and non-dry eye corneas (all P<0.05). The levels of RNA expression of VEGF-A, TNF-alpha, and IL-6 in the aflibercept subgroup were significantly decreased compared with those in the PBS subgroup in the dry eye group. CONCLUSION Compared with non-dry eye corneas, dry eye corneas have greater amounts of inflammation and neovascularization and also have a more robust response to anti-inflammatory and anti-angiogenic agents after ocular surface surgery. Trapping VEGF-A is effective in decreasing both angiogenesis and inflammation in dry eye corneas after ocular surface surgery. PMID:27990354

Preparation of acellular scaffold for corneal tissue engineering by supercritical carbon dioxide extraction technology.

PubMed

Huang, Yi-Hsun; Tseng, Fan-Wei; Chang, Wen-Hsin; Peng, I-Chen; Hsieh, Dar-Jen; Wu, Shu-Wei; Yeh, Ming-Long

2017-08-01

In this study, we developed a novel method using supercritical carbon dioxide (SCCO 2 ) to prepare acellular porcine cornea (APC). Under gentle extraction conditions using SCCO 2 technology, hematoxylin and eosin staining showed that cells were completely lysed, and cell debris, including nuclei, was efficiently removed from the porcine cornea. The SCCO 2 -treated corneas exhibited intact stromal structures and appropriate mechanical properties. Moreover, no immunological reactions and neovascularization were observed after lamellar keratoplasty in rabbits. All transplanted grafts and animals survived without complications. The transplanted APCs were opaque after the operation but became transparent within 2weeks. Complete re-epithelialization of the transplanted APCs was observed within 4weeks. In conclusion, APCs produced by SCCO 2 extraction technology could be an ideal and useful scaffold for corneal tissue engineering. We decellularized the porcine cornea using SCCO 2 extraction technology and investigated the characteristics, mechanical properties, and biocompatibility of the decellularized porcine cornea by lamellar keratoplasty in rabbits. To the best of our knowledge, this is the first report describing the use of SCCO 2 extraction technology for preparation of acellular corneal scaffold. We proved that the cellular components of porcine corneas had been efficiently removed, and the biomechanical properties of the scaffold were well preserved by SCCO 2 extraction technology. SCCO 2 -treated corneas maintained optical transparency and exhibited appropriate strength to withstand surgical procedures. In vivo, the transplanted corneas showed no evidence of immunological reactions and exhibited good biocompatibility and long-term stability. Our results suggested that the APCs developed by SCCO 2 extraction technology could be an ideal and useful scaffold for corneal replacement and corneal tissue engineering. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Rebamipide increases the amount of mucin-like substances on the conjunctiva and cornea in the N-acetylcysteine-treated in vivo model.

PubMed

Urashima, Hiroki; Okamoto, Takashi; Takeji, Yasuhiro; Shinohara, Hisashi; Fujisawa, Shigeki

2004-08-01

Rebamipide increases the amount of mucin-like substances in the stomach. We aimed to determine the effects of rebamipide on the amount of mucin-like substances in the conjunctiva and cornea of N-acetylcysteine-treated eyes. Furthermore, we attempted to evaluate the effects of rebamipide on the wound healing of N-acetylcysteine-treated eyes. The model was created by instilling 10% N-acetylcysteine solutions into rabbit eyes. Rebamipide was then applied on the day following the completion of N-acetylcysteine treatment. The amount of mucin-like substances on the conjunctiva and cornea was measured using the Alcian-blue binding method. The degree of damage was evaluated using scores based on the areas and densities of the cornea and conjunctival after staining using a rose Bengal solution under blind conditions. Rebamipide increased the level of mucin-like substances on the conjunctiva of N-acetylcysteine-treated eyes when instilled at concentrations of 0.3% or higher, and 1% rebamipide increased the amount of mucin-like substances covering the cornea. Moreover, 1% rebamipide improved the rose Bengal scores of the cornea and conjunctiva in N-acetylcysteine-treated eyes. Rebamipide increased mucin-like substances on the cornea and conjunctiva of N-acetylcysteine-treated eyes. In accordance with the mucin-increasing effects, rebamipide improved the rose Bengal scores for the cornea and conjunctiva of N-acetylcysteine-treated eyes. However, the relevance of these findings to dry eyes is unclear because it is not known whether the change in mucus expression in the N-acetylcysteine model is similar to what occurs in aqueous tear deficiency. Consequently, it may be worth trying on an animal model of keratoconjunctivitis sicca.

Temperature monitoring by infrared radiation measurements during ArF excimer laser ablation with cornea

NASA Astrophysics Data System (ADS)

Ishihara, Miya; Arai, Tsunenori; Sato, Shunichi; Nakano, Hironori; Obara, Minoru; Kikuchi, Makoto

1999-06-01

We measured infrared thermal radiation from porcine cornea during various fluences ArF excimer laser ablations with 1 microsecond(s) rise time. To obtain absolute temperature by means of Stefan-Boltzman law of radiation, we carried out a collection efficiency and detective sensitivity by a pre-experiment using panel heater. We measured the time course of the thermal radiation intensity with various laser fluences. We studied the relation between the peak cornea temperature during the ablation and irradiation fluences. We found the ablation situations, i.e., sub-ablation threshold, normal thermal ablation, and over-heated ablation, may be judged by both of the measured temperature transient waveforms and peak temperature. The boundary fluences corresponding to normal thermal ablation were 90 and 160 mJ/cm2. Our fast remote temperature monitoring during cornea ablation might be useful to control ablation quality/quantity of the cornea ArF laser ablation, that is PRK.

Optical anisotropy of the human cornea determined with a polarizing microscope.

PubMed

Bone, Richard A; Draper, Grenville

2007-12-01

We have investigated the optical anisotropy of the human cornea using a polarizing microscope normally used for optical mineralogy studies. The central part of the cornea was removed from 14 eyes (seven donors). With the sample placed on the microscope stage, we consistently observed hyperbolic isogyres characteristic of a negative biaxial material. The angle between the optic axes, generally similar in both eyes, ranged from 12 degrees to 40 degrees (mean+/-SD=31 degrees +/-8 degrees ). The optic axial plane always inclined downward in the nasal direction at 1 degrees -45 degrees below the horizontal (mean+/-SD=22+/-13 degrees ). The retardance produced by the corneas was estimated to be less than 200 nm. In conclusion, the human cornea possesses the anisotropy of a negative biaxial material. Both the angle between the optic axes and the retardance were fairly constant among the majority of samples, suggestive of uniformity in corneal structure.

Ultrastructure Features and Three-Dimensional Transmission Electron Tomography of Dhub Lizard (Uromastyx Aegyptia) Cornea and Its Adaptation to a Desert Environment.

PubMed

Akhtar, Saeed; Alkhalaf, Mousa; Khan, Adnan A; Almubrad, Turki M

2016-08-01

We report ultrastructural features and transmission electron tomography of the dhub lizard (Uromastyx aegyptia) cornea and its adaptation to hot and dry environments. Six corneas of dhub lizards were fixed in 2.5% glutaraldehyde and processed for electron microscopy and tomography. The ultrathin sections were observed with a JEOL 1400 transmission electron microscope. The cornea of the dhub lizard is very thin (~28-30 µm). The epithelium constitutes ~14% of the cornea, whereas the stroma constitutes 80% of the cornea. The middle stromal lamellae are significantly thicker than anterior and posterior stromal lamellae. Collagen fibril (CF) diameters in the anterior stroma are variable in size (25-75 nm). Proteoglycans (PGs) are very large in the middle and posterior stroma, whereas they are small in the anterior stroma. Three-dimensional electron tomography was carried out to understand the structure and arrangement of the PG and CFs. The presence of large PGs in the posterior and middle stroma might help the animal retain a large amount of water to protect it from dryness. The dhub corneal structure is equipped to adapt to the dry and hot desert environment.

Biocompatibility of poly(ethylene glycol) and poly(acrylic acid) interpenetrating network hydrogel by intrastromal implantation in rabbit cornea

PubMed Central

Zheng, Luo Luo; Vanchinathan, Vijay; Dalal, Roopa; Noolandi, Jaan; Waters, Dale J.; Hartmann, Laura; Cochran, Jennifer R.; Frank, Curtis W.; Yu, Charles Q.; Ta, Christopher N.

2015-01-01

We evaluated the biocompatibility of a poly(ethylene glycol) and poly(acrylic acid) (PEG/PAA) interpenetrating network hydrogel designed for artificial cornea in a rabbit model. PEG/PAA hydrogel measuring 6 mm in diameter was implanted in the corneal stroma of twelve rabbits. Stromal flaps were created with a microkeratome. Randomly, six rabbits were assigned to bear the implant for 2 months, two rabbits for 6 months, two rabbits for 9 months, one rabbit for 12 months, and one rabbit for 16 months. Rabbits were evaluated monthly. After the assigned period, eyes were enucleated, and corneas were processed for histology and immunohistochemistry. There were clear corneas in three of six rabbits that had implantation of hydrogel for 2 months. In the six rabbits with implant for 6 months or longer, the corneas remained clear in four. There was a high rate of epithelial defect and corneal thinning in these six rabbits. One planned 9-month rabbit developed extrusion of implant at 4 months. The cornea remained clear in the 16-month rabbit but histology revealed epithelial in-growth. Intrastromal implantation of PEG/PAA resulted in a high rate of long-term complications. PMID:25778285

Measuring effects of refractive surgery on corneas using Taylor series polynomials

NASA Astrophysics Data System (ADS)

Corbin, Jacob A.; Klein, Stanley A.; van de Pol, Corina

1999-06-01

Corneal topographers have made it possible to accurately map corneal shape. We applied this technology to model the post- refractive surgery cornea using Taylor series polynomials. Topography data was taken from 58 patient eyes with photorefractive keratectomy (PRK) or astigmatic photorefractive keratectomy (PARK). We looked at the changes the cornea underwent surgically, as well as the healing process. We compared the post-ablation cornea to the pre-ablation cornea and to the intended correction using novel topography maps. From the refractive map, we quantified the spherical aberration as areas of defocus on the cornea. From the pre-op exam to the first post-op exam, we measured 0.19+/-0.10 mm radius decrease in PRK and a 0.13+/-0.08 mm radius decrease in PARK in the areas where rays come to within two diopeters of defocus. As this change occurs within the optical zone, this corresponds to an increase in spherical aberration for both the PRK and the PARK patient. As the patient healed, we found additional decrease in radius of the zones of best vision in PRK patients, whereas we found no significant decrease in PARK patients. This corresponds to increased spherical aberration in the PRK patient.

Effects of nicergoline on corneal epithelial wound healing in rat eyes.

PubMed

Kim, Su-Young; Choi, Jun-Sub; Joo, Choun-Ki

2009-02-01

To investigate the effect of nicergoline on corneal epithelial wound healing in rats. One hundred Sprague-Dawley male rats were divided into two groups, the control group and the nicergoline-treated group, for 2 weeks. Corneal wound healing was evaluated by fluorescein staining after epithelial debridement. Nerve growth factor (NGF) protein and NGF mRNA were measured in rat corneas by ELISA and RT-PCR. NGF concentration of lacrimal gland was also evaluated by means of ELISA. Immunofluorescent staining was performed in rat corneas. The corneal wound healing rate was increased in nicergoline-treated rats compared with control rats after debridement. Twenty-four hours after epithelial debridement, corneal NGF protein and NGF mRNA levels were higher in the nicergoline-treated group than in the control group. Immunofluorescent staining showed that NGF staining was stronger in nicergoline-treated corneas than in control corneas 24 hours after epithelial debridement. In addition, NGF concentrations in lacrimal glands of the nicergoline-treated group were significantly higher than in the control group 24 hours after epithelial debridement. Nicergoline accelerated wound healing in rat corneas. The promoting effect of nicergoline in corneal wound healing is likely to be related to increased NGF in corneas and lacrimal glands.

Hypothermic preservation of corneas in a hyperkalaemic solution (CPTES): I. Short-term storage in the absence of colloid osmotic agents.

PubMed Central

Taylor, M J; Hunt, C J

1989-01-01

Preservation solutions for short-term storage of isolated donor corneas for use in penetrating keratoplasty have all been based on tissue culture medium, on the assumption that media designed to maintain the viability of cells at physiological temperatures will also provide suitable conditions for preservation at reduced temperatures. But for hypothermic preservation of some other tissues and organs, when ionic pumps are inhibited, it is unnecessary to support metabolism, and beneficial control of ion and water distribution between intra- and extracellular compartments is achieved by storage in appropriately formulated 'intracellular-type' solutions. We have therefore designed a solution that will restrict ionic imbalances and minimise endothelial cell swelling in corneas during exposure at reduced temperatures. This potassium-rich solution contains the biological pH buffer TES as an impermeant anion and is designated CPTES (corneal-potassium-TES). The structural and functional integrity of rabbit corneas stored at 0 degrees C in CPTES, without the addition of colloid osmotic agents, is compared with that of corneas stored in glutathione bicarbonate Ringers' solution (GBR), an 'extracellular-type' medium formulated for the maintenance of endothelial integrity during in-vitro perfusion at 34 degrees C. Corneas swelled significantly less during storage in CPTES than in GBR and could be stored for five days before reaching the same degree of hydration as corneas stored for only three days in GBR. Gross structural integrity and endothelial ultrastructure were maintained during storage for three and five days in CPTES. The rate of thinning of corneas stored in CPTES was significantly greater than in comparable groups of corneas stored in GBR. However, the efficient dehydration of corneas stored in CPTES was always preceded during perfusion by a brief period of additional swelling which was shown to be an osmotic response during the elution of the buffer compound TES that had permeated the stroma during storage. The omission of calcium or the addition of adenosine and glutathione to the CPTES preservation medium had no detectable effect on the integrity of the endothelium, but the omission of bicarbonate was beneficial, producing significantly higher rates of stromal thinning during normothermic perfusion. Additional benefits for extending storage by including colloid osmotic agents are described in a companion paper. Images PMID:2818988

Biomechanical and optical behavior of human corneas before and after photorefractive keratectomy.

PubMed

Sánchez, Paolo; Moutsouris, Kyros; Pandolfi, Anna

2014-06-01

To evaluate numerically the biomechanical and optical behavior of human corneas and quantitatively estimate the changes in refractive power and stress caused by photorefractive keratectomy (PRK). Athineum Refractive Center, Athens, Greece, and Politecnico di Milano, Milan, Italy. Retrospective comparative interventional cohort study. Corneal topographies of 10 human eyes were taken with a scanning-slit corneal topographer (Orbscan II) before and after PRK. Ten patient-specific finite element models were created to estimate the strain and stress fields in the cornea in preoperative and postoperative configurations. The biomechanical response in postoperative eyes was computed by directly modeling the postoperative geometry from the topographer and by reproducing the corneal ablation planned for the PRK with a numerical reprofiling procedure. Postoperative corneas were more compliant than preoperative corneas. In the optical zone, corneal thinning decreased the mechanical stiffness, causing local resteepening and making the central refractive power more sensitive to variations in intraocular pressure (IOP). At physiologic IOP, the postoperative corneas had a mean 7% forward increase in apical displacement and a mean 20% increase in the stress components at the center of the anterior surface over the preoperative condition. Patient-specific numerical models of the cornea can provide quantitative information on the changes in refractive power and in the stress field caused by refractive surgery. No author has a financial or proprietary interest in any material or method mentioned. Copyright © 2014 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

Effect of Intraocular Pressure and Anisotropy on the Optical Properties of the Cornea: A Study Using Polarization Sensitive Optical Coherence Tomography.

PubMed

Richhariya, Ashutosh; Verma, Yogesh; Rao, Divakar K; Roberts, Cynthia J; Mahmoud, Ashraf M; Sangwan, Virender S; Punjabi, Sunil; Gupta, Pradeep K

2014-01-01

We hypothesize that because of the anisotropic properties of the cornea, there should be a nonuniform change in birefringence with an increase in intraocular pressure (IOP). In this in vitro study, anisotropic properties, stress distribution within the cornea, and the effect of IOP on changes in stress level were investigated. Button inflation tests for deformation with polarization sensitive optical coherence tomography were used to demonstrate optical and material anisotropy on ex vivo human corneas. Inflation tests were performed on human donor corneoscleral rims. Using a turntable and hydrostatic column, each corneoscleral rim was subjected to a hydrostatic pressure of 0, 10, 15, and 20 mm Hg. At each pressure step, 4 scans at 0, 45, 90, and 135 degrees were taken by a polarization sensitive optical coherence tomography system, and the birefringence images and normal intensity-based images were recorded; images were later compiled for analysis. The retardation changed with the axis of orientation (P [T ≤ t] 1-tailed = 0.025) and IOP (P [T ≤ t] 1-tailed = 0.019). Optical thickness of the cornea decreased with increasing IOP. The optical properties of the cornea are modified with change in IOP. This is not uniform because of distinct anisotropic properties. Anisotropic properties may unpredictably affect the optical quality of cornea during or after the surgeries. Changes in corneal birefringence can be also used as a tool for measuring the IOP of the eye.

Limited Versus Total Epithelial Debridement Ocular Surface Injury: Live Fluorescence Imaging of Hemangiogenesis and Lymphangiogenesis in Prox1-GFP/Flk1::myr-mCherry Mice

PubMed Central

Chang, Jin-Hong; Putra, Ilham; Huang, Yu-hui; Chang, Michael; Han, Kyuyeon; Zhong, Wei; Gao, Xinbo; Wang, Shuangyong; Dugas-Ford, Jennifer; Nguyen, Tara; Hong, Young-Kwon; Azar, Dimitri T.

2016-01-01

Background Immunohistochemical staining experiments have shown that both hemangiogenesis and lymphangiogenesis occur following severe corneal and conjunctival injury and that the neovascularization of the cornea often has severe visual consequences. To better understand how hemangiogenesis and lymphangiogenesis are induced by different degrees of ocular injury, we investigated patterns of injury-induced corneal neovascularization in live Prox1-GFP/Flk1::myr-mCherry mice, in which blood and lymphatic vessels can be imaged simultaneously in vivo. Methods The eyes of Prox1-GFP/Flk1::myr-mCherry mice were injured according to four models based on epithelial debridement of the: A) central cornea (a 1.5-mm-diameter circle of tissue over the corneal apex), B) total cornea, C) bulbar conjunctiva, and D) cornea+bulbar conjunctiva. Corneal blood and lymphatic vessels were imaged on days 0, 3, 7, and 10 post-injury, and the percentages of the cornea containing blood and lymphatic vessels were calculated. Results Neither central corneal nor bulbar conjunctival debridement resulted in significant vessel growth in the mouse cornea, whereas total corneal and corneal+bulbar conjunctival debridement did. On day 10 in the central cornea, total cornea, bulbar conjunctiva, and corneal+bulbar conjunctival epithelial debridement models, the percentage of the corneal surface that was occupied by blood vessels (hemangiogenesis) was 1.9±0.8%, 7.14±2.4%, 2.29±1%, and 15.05±2.14%, respectively, and the percentage of the corneal surface that was occupied by lymphatic vessels (lymphangiogenesis) was 2.45±1.51%, 4.85±0.95%, 2.95±1.27%, and 4.15±3.85%, respectively. Conclusions Substantial corneal debridement was required to induce corneal neovascularization in the mouse cornea, and the corneal epithelium may therefore be partially responsible for maintaining corneal avascularity. General significance Our study demonstrates that GFP/Flk1::myr-mCherry mice are a useful model for studying coordinated hemangiogenic and lymphangiogenic responses. PMID:27233452

[The transfection and expression of IL-1ra gene to the rabbit cornea in situ via cation polymer mediation].

PubMed

Yuan, Jin; Chen, Jia-qi; Zhou, Shi-you; Liu, Zu-guo; Wang, Zhi-chong; Gu, Jian-jun

2006-08-01

To investigate the efficiency and safety of transfection of PEGFP-IL-1ra plasmid via cation polymer mediation (poly-ethylenimine, PEI) by injection into the corneal stroma. Human IL-1ra cDNA fragments were cloned by RT-PCR. Plasmid PEGFP-hIL-1ra recombinants were constructed and transferred into corneal endothelial cells (CEC) via cation polymer mediation. Expression of IL-1ra mRNA and IL-1ra was detected by green fluorescent protein (GFP) and Western-blotting. In the experiment group, 20 microl preparation containing 10 microg plasmid PEGFP-hIL-1ra recombinants and PEI-in-vivo was injected into the corneal stroma of Wistar rats (n = 30). Equivalent PEI-in-vivo solution was injected into another 15 corneas as the controls. Corneas were harvested at different time points (day 1, 3, 6, 14 and 21) after injection. The changes of tissue structure and function after IL-1ra in situ transfection were studied by HE staining, transmission electron microscopy, trypan blue-alizarin red staining and immunohistochemistry. The location and intensity of IL-1ra-GFP fusion protein expression were monitored by fluorescence microscopy. The size of the RT-PCR product of hIL-1ra fragments was approximately 500 bp in agarose gel electrophoresis. Restrictive enzyme digestion analysis of PstI, BamHI and DNA sequence analysis showed that expression of plasmid PEGFP-hIL-1ra recombinants had been constructed successfully. Twelve hours after the transfection of PEGFP-hIL-1ra, GFP fluorescence was detected in 10% - 15% endothelial cells. IL-1ra protein (RMW: 44,000) was detected by Western-blotting. In PEGFP-hIL-1ra treated group, fluorescence was appeared at day 1 in cornea basal epithelial cells, peaked at day 6 in whole cornea, began to weaken at day 14, and only weak fluorescence remained in cornea epithelial cells at day 21. No fluorescence appeared in the control group. No significant pathologic changes could be found in HE stained cornea tissues in both transfected group and the controls. p63 immunocytochemical staining in cornea epithelium was positive in both groups. Trypan blue-alizarin red staining confirmed that there was no damage in cornea endothelial cells. IL-1ra-GFP granules could be found by transmission electron microscope in every layer of cornea in the transfected group, but none in the controls. There was no impairment in the ultrastructure of cells in both groups. By direct injection of PEGFP-hIL-1ra into corneal stroma and mediated by cation polymer, IL-1ra genes could be transferred and expressed in corneal tissue efficiently and safely, and might provide a novel technique of gene transfection to cornea in situ.

Lens regeneration from the cornea requires suppression of Wnt/β-catenin signaling.

PubMed

Hamilton, Paul W; Sun, Yu; Henry, Jonathan J

2016-04-01

The frog, Xenopus laevis, possesses a high capacity to regenerate various larval tissues, including the lens, which is capable of complete regeneration from the cornea epithelium. However, the molecular signaling mechanisms of cornea-lens regeneration are not fully understood. Previous work has implicated the involvement of the Wnt signaling pathway, but molecular studies have been very limited. Iris-derived lens regeneration in the newt (Wolffian lens regeneration) has shown a necessity for active Wnt signaling in order to regenerate a new lens. Here we provide evidence that the Wnt signaling pathway plays a different role in the context of cornea-lens regeneration in Xenopus. We examined the expression of frizzled receptors and wnt ligands in the frog cornea epithelium. Numerous frizzled receptors (fzd1, fzd2, fzd3, fzd4, fzd6, fzd7, fzd8, and fzd10) and wnt ligands (wnt2b.a, wnt3a, wnt4, wnt5a, wnt5b, wnt6, wnt7b, wnt10a, wnt11, and wnt11b) are expressed in the cornea epithelium, demonstrating that this tissue is transcribing many of the ligands and receptors of the Wnt signaling pathway. When compared to flank epithelium, which is lens regeneration incompetent, only wnt11 and wnt11b are different (present only in the cornea epithelium), identifying them as potential regulators of cornea-lens regeneration. To detect changes in canonical Wnt/β-catenin signaling occurring within the cornea epithelium, axin2 expression was measured over the course of regeneration. axin2 is a well-established reporter of active Wnt/β-catenin signaling, and its expression shows a significant decrease at 24 h post-lentectomy. This decrease recovers to normal endogenous levels by 48 h. To test whether this signaling decrease was necessary for lens regeneration to occur, regenerating eyes were treated with either 6-bromoindirubin-3'-oxime (BIO) or 1-azakenpaullone - both activators of Wnt signaling - resulting in a significant reduction in the percentage of cases with successful regeneration. In contrast, inhibition of Wnt signaling using either the small molecule IWR-1, treatment with recombinant human Dickkopf-1 (rhDKK1) protein, or transgenic expression of Xenopus DKK1, did not significantly affect the percentage of successful regeneration. Together, these results suggest a model where Wnt/β-catenin signaling is active in the cornea epithelium and needs to be suppressed during early lens regeneration in order for these cornea cells to give rise to a new lentoid. While this finding differs from what has been described in the newt, it closely resembles the role of Wnt signaling during the initial formation of the lens placode from the surface ectoderm during early embryogenesis. Copyright © 2016 Elsevier Ltd. All rights reserved.

Optics of the human cornea influence the accuracy of stereo eye-tracking methods: a simulation study.

PubMed

Barsingerhorn, A D; Boonstra, F N; Goossens, H H L M

2017-02-01

Current stereo eye-tracking methods model the cornea as a sphere with one refractive surface. However, the human cornea is slightly aspheric and has two refractive surfaces. Here we used ray-tracing and the Navarro eye-model to study how these optical properties affect the accuracy of different stereo eye-tracking methods. We found that pupil size, gaze direction and head position all influence the reconstruction of gaze. Resulting errors range between ± 1.0 degrees at best. This shows that stereo eye-tracking may be an option if reliable calibration is not possible, but the applied eye-model should account for the actual optics of the cornea.

BALB/c and C57BL6 mouse strains vary in their ability to heal corneal epithelial debridement wounds

PubMed Central

Pal-Ghosh, Sonali; Tadvalkar, Gauri; Jurjus, Rosalyn A.; Zieske, James D.; Stepp, Mary Ann

2008-01-01

Genetically engineered mice are usually produced on a mixed genetic background and can be derived from several mouse strains including 129SvJ, C57BL6, and BALB/c. To determine whether differences in recurrent corneal epithelial erosions (RCEEs), corneal epithelial stem cell deficiency (CESCD), and cell migration rate vary between two different mouse strains (BALB/c and C57BL6), 8 week mice were subjected to 1.5 (small) or 2.8 mm (large) manual debridement wounds and allowed to heal for 4 weeks. Syndecan-1 (sdc-1) null mice backcrossed seven generations onto a BALB/c genetic background were also included in the RCEE and CESCD studies to permit comparisons between genotypes within a single strain. After sacrifice, corneas were assessed for the presence of recurrent erosions; no fewer than 15 corneas were used for each strain or genotype studied. Data show that the frequency of recurrent erosions after small wounds was 81 +/− 9% in the C57BL6 mice, 73 +/− 2% in the BALB/c mice, and 32 +/− 6% in sdc-1 null mice. Neither strain developed CESCD after small wounds. The frequency of erosions after large wounds was greater (88 +/− 8%) in the C57BL6 mice compared to BALB/c (60 +/− 2%), and sdc-1 null mice (32 +/− 5%). 4 weeks after the large wounds, fixed, flat mounted corneas were assessed for evidence of CESCD with antibodies against the conjunctival keratin K8 and the goblet cell marker, the mucin Muc5AC. The frequency of CESCD 4 weeks after the large wounds was significantly greater in the C57BL6 mice than in the BALB/c or sdc-1 null mice. To assess cell migration rates, corneas were subjected to 1.5 mm wounds and allowed to heal for 12, 15, 18, 21, and 24 hours. After sacrifice, corneas were stained with Richardson stain (BALB/c) or propidium iodide (C57BL6) to assess reepithelialization rates. While reepithelialization rates were similar for the early times after wounding, by 24 hours the C57BL6 corneas had healed faster: 16 of 30 corneas from the C57BL6 mice were closed compared to 9 of 30 of the BALB/c wounds. BALB/c corneas appeared larger overall compared to C57BL6 corneas; measurements of the overall mass of the enucleated eyes and diameters of the flat-mounted corneas confirmed that C57BL6 eyes and corneas were 6.8% and 4.4% smaller respectively than those of BALB/c mice even though the masses of the two mouse strains at 8 weeks of age were identical. Using BrdU to label dividing cells, we found that 18 hours after wounding, C57BL6 and BALB/c corneal epithelia showed similar numbers of proliferating cells. To determine if the enhanced corneal epithelial cell migration rate seen in the C57BL6 mice was specific to the cornea, we conducted time-lapse studies to assess random cell migration rates in vitro using primary cultures of mouse epidermal keratinocytes. Consistent with the in vivo data, epidermal keratinocytes derived from BALB/c mice migrated 60% slower than C57BL6 cells. These data prove that strain-specific differences in cell migration rate in vivo are present in the cornea and are accompanied by differences in the frequencies of recurrent erosions and corneal epithelial stem cell deficiency. PMID:18809399

Donor cornea preparation in partial big bubble deep anterior lamellar keratoplasty.

PubMed

Lim, Li; Lim, Samuel Wen Yan

2014-01-01

The purpose of this paper is to describe a technique of donor cornea preparation to ensure good graft-host apposition in incomplete big bubble deep anterior lamellar keratoplasty. Following a partial-thickness trephination, manual dissection and excision of corneal stroma was performed. Anwar's big-bubble technique involving a deep stromal air injection was then initiated. However, the big bubble could not extend to the trephination edge and the peripheral residual corneal stroma could not be removed. Donor cornea preparation involving trimming of the posterior lip of the corneal button was then performed and good graft-host apposition was obtained without graft over-ride. We performed peripheral donor cornea trimming prior to allograft placement in order to ensure good graft-host apposition. Postoperatively, best-corrected visual acuity in both eyes was 6/7.5. Donor cornea preparation involving trimming of the posterior lip of the corneal button is a useful technique in instances where the big bubble does not extend to the trephination edge and ensures good graft-host apposition.

THz imaging system for in vivo human cornea.

PubMed

Sung, Shijun; Selvin, Skyler; Bajwa, Neha; Chantra, Somporn; Nowroozi, Bryan; Garritano, James; Goell, Jacob; Li, Alex; Deng, Sophie X; Brown, Elliott; Grundfest, Warren S; Taylor, Zachary D

2018-01-01

Terahertz (THz) imaging of corneal tissue water content (CTWC) is a proposed method for early, accurate detection and study of corneal diseases. Despite promising results from ex vivo and in vivo cornea studies, interpretation of the reflectivity data is confounded by the contact between corneal tissue and rigid dielectric window used to flatten the imaging field. This work develops a novel imaging system and image reconstruction methods specifically for nearly spherical targets such as human cornea. A prototype system was constructed using a 650 GHz multiplier source and Schottky diode detector. Resolution and imaging field strength measurement from characterization targets correlate well with those predicted by the quasioptical theory and physical optics analysis. Imaging experiments with corneal phantoms and ex vivo corneas demonstrate the hydration sensitivity of the imaging system and reliable measurement of CTWC. We present successful acquisition of non-contact THz images of in vivo human cornea, and discuss strategies for optimizing the imaging system design for clinical use.

Photoablation of the cornea with a Q-switched Er:YAG laser

NASA Astrophysics Data System (ADS)

Lubatschowski, Holger; Hetzel, U.; Kermani, Omid; Ziolek, Carsten; Drommer, Wolfgang; Ertmer, Wolfgang

1997-12-01

In this study the ablation characteristics and the wound healing process of rabbit cornea irradiated with a Q- switched Er:YAG laser was evaluated. The laser, emitting at 2.94 micrometers wavelength, has a pulse width of 100 ns. The spot size on the corneal surface was 1 mm in diameter at a fluence of 750 mJ/cm2. The laser beam was applied by a `flying spot' mode, performing refractive ablations of -7 to -8 dpt. As a biological model, the corneas of 9 rabbits were irradiated. The post-treatment follow-up was as long as 39 days. The treated corneas were investigated by light and electron microscopy. The wound healing on rabbit cornea of the Q-switched Er:YAG laser radiation in corneal tissue processing resembles to what is known from ArF- excimer laser application. To shorten the pulse width by means of Q-switching is one major key to the successful application of the Er:YAG laser for PRK.

Interfaces detection after corneal refractive surgery by low coherence optical interferometry

PubMed Central

Verrier, I.; Veillas, C.; Lépine, T.; Nguyen, F.; Thuret, G.; Gain, P.

2010-01-01

The detection of refractive corneal surgery by LASIK, during the storage of corneas in Eye Banks will become a challenge when the numerous operated patients will arrive at the age of cornea donation. The subtle changes of corneal structure and refraction are highly suspected to negatively influence clinical results in recipients of such corneas. In order to detect LASIK cornea interfaces we developed a low coherence interferometry technique using a broadband continuum source. Real time signal recording, without moving any optical elements and without need of a Fourier Transform operation, combined with good measurement resolution is the main asset of this interferometer. The associated numerical processing is based on a method initially used in astronomy and offers an optimal correlation signal without the necessity to image the whole cornea that is time consuming. The detection of corneal interfaces - both outer and inner surface and the buried interface corresponding to the surgical wound – is then achieved directly by the innovative combination of interferometry and this original numerical process. PMID:21258562

Impaired Fas-Fas Ligand Interactions Result in Greater Recurrent Herpetic Stromal Keratitis in Mice

PubMed Central

Yin, Xiao-Tang; Keadle, Tammie L.; Hard, Jessicah; Herndon, John; Potter, Chloe A.; Del Rosso, Chelsea R.; Ferguson, Thomas A.; Stuart, Patrick M.

2015-01-01

Herpes simplex virus-1 (HSV-1) infection of the cornea leads to a potentially blinding condition termed herpetic stromal keratitis (HSK). Clinical studies have indicated that disease is primarily associated with recurrent HSK following reactivation of a latent viral infection of the trigeminal ganglia. One of the key factors that limit inflammation of the cornea is the expression of Fas ligand (FasL). We demonstrate that infection of the cornea with HSV-1 results in increased functional expression of FasL and that mice expressing mutations in Fas (lpr) and FasL (gld) display increased recurrent HSK following reactivation compared to wild-type mice. Furthermore, both gld and lpr mice took longer to clear their corneas of infectious virus and the reactivation rate for these strains was significantly greater than that seen with wild-type mice. Collectively, these findings indicate that the interaction of Fas with FasL in the cornea restricts the development of recurrent HSK. PMID:26504854

Impaired Fas-Fas Ligand Interactions Result in Greater Recurrent Herpetic Stromal Keratitis in Mice.

PubMed

Yin, Xiao-Tang; Keadle, Tammie L; Hard, Jessicah; Herndon, John; Potter, Chloe A; Del Rosso, Chelsea R; Ferguson, Thomas A; Stuart, Patrick M

2015-01-01

Herpes simplex virus-1 (HSV-1) infection of the cornea leads to a potentially blinding condition termed herpetic stromal keratitis (HSK). Clinical studies have indicated that disease is primarily associated with recurrent HSK following reactivation of a latent viral infection of the trigeminal ganglia. One of the key factors that limit inflammation of the cornea is the expression of Fas ligand (FasL). We demonstrate that infection of the cornea with HSV-1 results in increased functional expression of FasL and that mice expressing mutations in Fas (lpr) and FasL (gld) display increased recurrent HSK following reactivation compared to wild-type mice. Furthermore, both gld and lpr mice took longer to clear their corneas of infectious virus and the reactivation rate for these strains was significantly greater than that seen with wild-type mice. Collectively, these findings indicate that the interaction of Fas with FasL in the cornea restricts the development of recurrent HSK.

Effect of topical antiinflammatory drugs on mechanical behavior of rabbit cornea.

PubMed

Lepore, Domenico; De Santis, Roberto; Pagliara, Monica M; Gloria, Antonio; Oliviero, Olimpia; Nucci, Carlo; Improta, Giovanni; Triassi, Maria; Ambrosio, Luigi

2017-04-26

A variety of antiinflammatory therapies are employed to promote corneal wound healing. The effects of steroidal and nonsteroidal antiinflammatory drugs on the biomechanical properties of rabbit cornea were investigated over time using tensile tests. Full-thickness incisions were made and used to analyze the effects of dexamethasone sodium phosphate 0.1% and diclofenac sodium 0.1% on corneal biomechanical properties during wound healing at 7, 14 and 21 days after surgery. The full-thickness incision deeply modified all of the mechanical properties. At 3 weeks after incision, regardless of the drug therapy, the tensile modulus was about 70% of the value for the intact cornea. Topical treatment with dexamethasone was particularly effective during the first week after surgery; the second week after surgery, a similar result was observed in the corneas treated with diclofenac. Low doses of steroidal and nonsteroidal antiinflammatory drugs would seem to have the potential to improve biomechanical properties only during the early stage of the healing process of the cornea.

Inflammation Modulatory Protein TSG-6 for Chemical Injuries to the Cornea

DTIC Science & Technology

2016-10-01

presented February 2016 at the World Ophthalmology Congress in Guadalajara Mexico titled: Comprehensive Profiling of Alkali Injuries to the Cornea. Appendix...World Ophthalmology Congress in February 2016. If we are able to establish efficacy of IV TSG-6 in this model, it would provide the first effective...Publications/papers/presentations 1) Paper presentation Comprehensive Profiling of Alkali Injuries to the Cornea, presented at the World Ophthalmology

Development of a reconstructed cornea from collagen-chondroitin sulfate foams and human cell cultures.

PubMed

Vrana, N Engin; Builles, Nicolas; Justin, Virginie; Bednarz, Jurgen; Pellegrini, Graziella; Ferrari, Barbara; Damour, Odile; Hulmes, David J S; Hasirci, Vasif

2008-12-01

To develop an artificial cornea, the ability to coculture the different cell types present in the cornea is essential. Here the goal was to develop a full-thickness artificial cornea using an optimized collagen-chondroitin sulfate foam, with a thickness close to that of human cornea, by coculturing human corneal epithelial and stromal cells and transfected human endothelial cells. Corneal extracellular matrix was simulated by a porous collagen/glycosaminoglycan-based scaffold seeded with stromal keratocytes and then, successively, epithelial and endothelial cells. Scaffolds were characterized for bulk porosity and pore size distribution. The performance of the three-dimensional construct was studied by histology, immunofluorescence, and immunohistochemistry. The scaffold had 85% porosity and an average pore size of 62.1 microm. Keratocytes populated the scaffold and produced a newly synthesized extracellular matrix as characterized by immunohistochemistry. Even though the keratocytes lost their CD34 phenotype marker, the absence of smooth muscle actin fibers showed that these cells had not differentiated into myofibroblasts. The epithelial cells formed a stratified epithelium and began basement membrane deposition. An endothelial cell monolayer beneath the foam was also apparent. These results demonstrate that collagen-chondroitin sulfate scaffolds are good substrates for artificial cornea construction with good resilience, long-term culture capability, and handling properties.

Optical coherence tomography analysis of hydrofluoric acid decontamination of human cornea by mannitol solution.

PubMed

Nosé, Ricardo M; Daga, Fabio B; Nosé, Walton; Kasahara, Niro

2017-03-01

To evaluate the efficacy of mannitol solution as a decontamination agent on the chemical burn of the human corneas. Eight donor corneas from an eye bank were exposed to 25μl of 2.5% hydrofluoric acid (HF) solution on a filter paper for 20s. Three eyes were rinsed with 1000ml of mannitol 20% for 15min immediately after removal of the filter paper, 3 other were rinsed with sodium chloride (NaCl) 0.9% (1000ml for 15min) and two eyes were not rinsed. Microstructural changes were monitored in the time domain by optical coherence tomography (OCT) imaging for 75min. NaCl reduced the penetration depth to approximately half the thickness of the cornea at 15min; scattering within the anterior cornea was higher than that for the unrinsed eye. With mannitol, no increased scattering was observed in the posterior part of the corneal stroma within a time period of 1h after rinsing. OCT images revealed low-scattering intensity within the anterior stroma at the end of the rinsing period. In eye bank human corneas, mannitol proved to be an efficient agent to decontaminate HF burn. Copyright © 2016 Elsevier Ltd and ISBI. All rights reserved.

AAV Gene Therapy for MPS1-associated Corneal Blindness.

PubMed

Vance, Melisa; Llanga, Telmo; Bennett, Will; Woodard, Kenton; Murlidharan, Giridhar; Chungfat, Neil; Asokan, Aravind; Gilger, Brian; Kurtzberg, Joanne; Samulski, R Jude; Hirsch, Matthew L

2016-02-22

Although cord blood transplantation has significantly extended the lifespan of mucopolysaccharidosis type 1 (MPS1) patients, over 95% manifest cornea clouding with about 50% progressing to blindness. As corneal transplants are met with high rejection rates in MPS1 children, there remains no treatment to prevent blindness or restore vision in MPS1 children. Since MPS1 is caused by mutations in idua, which encodes alpha-L-iduronidase, a gene addition strategy to prevent, and potentially reverse, MPS1-associated corneal blindness was investigated. Initially, a codon optimized idua cDNA expression cassette (opt-IDUA) was validated for IDUA production and function following adeno-associated virus (AAV) vector transduction of MPS1 patient fibroblasts. Then, an AAV serotype evaluation in human cornea explants identified an AAV8 and 9 chimeric capsid (8G9) as most efficient for transduction. AAV8G9-opt-IDUA administered to human corneas via intrastromal injection demonstrated widespread transduction, which included cells that naturally produce IDUA, and resulted in a >10-fold supraphysiological increase in IDUA activity. No significant apoptosis related to AAV vectors or IDUA was observed under any conditions in both human corneas and MPS1 patient fibroblasts. The collective preclinical data demonstrate safe and efficient IDUA delivery to human corneas, which may prevent and potentially reverse MPS1-associated cornea blindness.

AAV Gene Therapy for MPS1-associated Corneal Blindness

PubMed Central

Vance, Melisa; Llanga, Telmo; Bennett, Will; Woodard, Kenton; Murlidharan, Giridhar; Chungfat, Neil; Asokan, Aravind; Gilger, Brian; Kurtzberg, Joanne; Samulski, R. Jude; Hirsch, Matthew L.

2016-01-01

Although cord blood transplantation has significantly extended the lifespan of mucopolysaccharidosis type 1 (MPS1) patients, over 95% manifest cornea clouding with about 50% progressing to blindness. As corneal transplants are met with high rejection rates in MPS1 children, there remains no treatment to prevent blindness or restore vision in MPS1 children. Since MPS1 is caused by mutations in idua, which encodes alpha-L-iduronidase, a gene addition strategy to prevent, and potentially reverse, MPS1-associated corneal blindness was investigated. Initially, a codon optimized idua cDNA expression cassette (opt-IDUA) was validated for IDUA production and function following adeno-associated virus (AAV) vector transduction of MPS1 patient fibroblasts. Then, an AAV serotype evaluation in human cornea explants identified an AAV8 and 9 chimeric capsid (8G9) as most efficient for transduction. AAV8G9-opt-IDUA administered to human corneas via intrastromal injection demonstrated widespread transduction, which included cells that naturally produce IDUA, and resulted in a >10-fold supraphysiological increase in IDUA activity. No significant apoptosis related to AAV vectors or IDUA was observed under any conditions in both human corneas and MPS1 patient fibroblasts. The collective preclinical data demonstrate safe and efficient IDUA delivery to human corneas, which may prevent and potentially reverse MPS1-associated cornea blindness. PMID:26899286

Prevalidation of a human cornea construct as an alternative to animal corneas for in vitro drug absorption studies.

PubMed

Hahne, Matthias; Zorn-Kruppa, Michaela; Guzman, Gustavo; Brandner, Johanna M; Haltner-Ukomado, Eleonore; Wätzig, Hermann; Reichl, Stephan

2012-08-01

The use of ophthalmic drugs has increased consistently over the past few decades. Currently, most research is conducted using in vivo and ex vivo animal experiments; however, they have many disadvantages, including ethical concerns, high costs, the questionable extension of animal results to humans, and poor standardization. Although several cell culture-based cornea models have been developed, none have been validated and accepted for general use. In this study, a standardized, three-dimensional model of the human cornea (Hemicornea, HC) based on immortalized human corneal cells and cultivated in serum-free conditions was developed for drug absorption studies and prevalidated using compounds with a wide range of molecular characteristics (sodium fluorescein, rhodamine B, fluorescein isothiocyanate-labeled dextran, aciclovir, bimatoprost, dexamethasone, and timolol maleate). The HC model was independently cultured in three different laboratories, and the intralaboratory and interlaboratory reproducibility was analyzed and compared with the rabbit cornea. This analysis showed that the HC has a barrier in the same range as excised animal corneas, although with a higher reproducibility and lower variability. Because of the demonstrated transferability, the HC represents a promising in vitro alternative to the use of ex vivo tissue and offers a well-defined and standardized system for drug absorption studies. Copyright © 2012 Wiley Periodicals, Inc.

Cornea nerve fiber quantification and construction of phenotypes in patients with fibromyalgia

PubMed Central

Oudejans, Linda; He, Xuan; Niesters, Marieke; Dahan, Albert; Brines, Michael; van Velzen, Monique

2016-01-01

Cornea confocal microscopy (CCM) is a novel non-invasive method to detect small nerve fiber pathology. CCM generally correlates with outcomes of skin biopsies in patients with small fiber pathology. The aim of this study was to quantify the morphology of small nerve fibers of the cornea of patients with fibromyalgia in terms of density, length and branching and further phenotype these patients using standardized quantitative sensory testing (QST). Small fiber pathology was detected in the cornea of 51% of patients: nerve fiber length was significantly decreased in 44% of patients compared to age- and sex-matched reference values; nerve fiber density and branching were significantly decreased in 10% and 28% of patients. The combination of the CCM parameters and sensory tests for central sensitization, (cold pain threshold, mechanical pain threshold, mechanical pain sensitivity, allodynia and/or windup), yielded four phenotypes of fibromyalgia patients in a subgroup analysis: one group with normal cornea morphology without and with signs of central sensitization, and a group with abnormal cornea morphology parameters without and with signs of central sensitization. In conclusion, half of the tested fibromyalgia population demonstrates signs of small fiber pathology as measured by CCM. The four distinct phenotypes suggest possible differences in disease mechanisms and may require different treatment approaches. PMID:27006259

In Vivo Confocal Microscopic Evaluation of Corneal Langerhans Cells in Dry Eye Patients§

PubMed Central

Machetta, Federica; Fea, Antonio M; Actis, Alessandro G; de Sanctis, Ugo; Dalmasso, Paola; Grignolo, Federico M

2014-01-01

Purpose. To assess inflammatory involvement of cornea in dry eye by means of confocal microscopy, evaluating the presence and distribution of Langherans cells (LCs). Methods: 98 eyes of 49 subjects were enrolled: 18 subjects affected by Sjögren Syndrome Dry Eye (SSDE), 17 with Non-Sjögren Syndrome Dry Eye (NSSDE), 14 healthy volunteeers. Dry eye symptoms, tear film, ocular surface damage and corneal confocal microscopy were analized. Results: A significant increase of LCs density was observed at sub-basal nerve plexus (SSDE = 79 cells/mm2 andNDE = 22 cells/mm2; p = 0,0031) and sub-epithelial nerve plexus (SSDE = 38 cells/mm2 and NDE = 3 cells/mm2; p = 0,0169) in central cornea of SSDE group. An increased number of LCs from the center to the periphery of the cornea was observed, significant only in healthy volunteers group. In dry eye patients there was an increase in LCs density in both peripheral and central cornea with a significant difference between NDE (14,66 cells/mm2) and SSDE (56,66 cells/mm2) only in central cornea (p = 0,0028). In SSDE group, mean density of LCs in central cornea results also superior to NSSDE group (29,33 cells/mm2). There was no correlation between LCs density and dry eye symptoms, tear film deficiency and ocular surface damage. Conclusion: This study demonstrates the activation of an inflammatory and immunological reaction in cornea of NSSDE and SSDE patients. Confocal microscopy can be an important diagnostic tool in evaluation and follow-up of dry eye disease. PMID:25317216

[Porous matrix and primary-cell culture: a shared concept for skin and cornea tissue engineering].

PubMed

Auxenfans, C; Builles, N; Andre, V; Lequeux, C; Fievet, A; Rose, S; Braye, F-M; Fradette, J; Janin-Manificat, H; Nataf, S; Burillon, C; Damour, O

2009-06-01

Skin and cornea both feature an epithelium firmly anchored to its underlying connective compartment: dermis for skin and stroma for cornea. A breakthrough in tissue engineering occurred in 1975 when skin stem cells were successfully amplified in culture by Rheinwald and Green. Since 1981, they are used in the clinical arena as cultured epidermal autografts for the treatment of patients with extensive burns. A similar technique has been later adapted to the amplification of limbal-epithelial cells. The basal layer of the limbal epithelium is located in a transitional zone between the cornea and the conjunctiva and contains the stem cell population of the corneal epithelium called limbal-stem cells (LSC). These cells maintain the proper renewal of the corneal epithelium by generating transit-amplifying cells that migrate from the basal layer of the limbus towards the basal layer of the cornea. Tissue-engineering protocols enable the reconstruction of three-dimensional (3D) complex tissues comprising both an epithelium and its underlying connective tissue. Our in vitro reconstruction model is based on the combined use of cells and of a natural collagen-based biodegradable polymer to produce the connective-tissue compartment. This porous substrate acts as a scaffold for fibroblasts, thereby, producing a living dermal/stromal equivalent, which once epithelialized results into a reconstructed skin/hemicornea. This paper presents the reconstruction of surface epithelia for the treatment of pathological conditions of skin and cornea and the development of 3D tissue-engineered substitutes based on a collagen-GAG-chitosan matrix for the regeneration of skin and cornea.

Three-dimensional confocal microscopy of the living cornea and ocular lens

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1991-07-01

The three-dimensional reconstruction of the optic zone of the cornea and the ocular crystalline lens has been accomplished using confocal microscopy and volume rendering computer techniques. A laser scanning confocal microscope was used in the reflected light mode to obtain the two-dimensional images from the cornea and the ocular lens of a freshly enucleated rabbit eye. The light source was an argon ion laser with a 488 nm wavelength. The microscope objective was a Leitz X25, NA 0.6 water immersion lens. The 400 micron thick cornea was optically sectioned into 133 three micron sections. The semi-transparent cornea and the in-situ ocular lens was visualized as high resolution, high contrast two-dimensional images. The structures observed in the cornea include: superficial epithelial cells and their nuclei, basal epithelial cells and their 'beaded' cell borders, basal lamina, nerve plexus, nerve fibers, nuclei of stromal keratocytes, and endothelial cells. The structures observed in the in- situ ocular lens include: lens capsule, lens epithelial cells, and individual lens fibers. The three-dimensional data sets of the cornea and the ocular lens were reconstructed in the computer using volume rendering techniques. Stereo pairs were also created of the two- dimensional ocular images for visualization. The stack of two-dimensional images was reconstructed into a three-dimensional object using volume rendering techniques. This demonstration of the three-dimensional visualization of the intact, enucleated eye provides an important step toward quantitative three-dimensional morphometry of the eye. The important aspects of three-dimensional reconstruction are discussed.

Influence of glucocorticosteroids on the biomechanical properties of in-vivo rabbit cornea.

PubMed

Yu, Ji-guo; Bao, Fang-jun; Joda, Akram; Fu, Xun-an; Zhou, Shi; Wang, Jing; Hu, Xiu-li; Wang, Qin-mei; Elsheikh, Ahmed

2014-01-01

Understanding corneal biomechanical responses during long-term glucocorticosteroids administration is important in clinical practice. The purpose of this study is to investigate the biomechanical influence of fluorometholone 0.1% eye drops on rabbit cornea. Thirty-eight Japanese white rabbits were randomly divided into three groups; a fluorometholone group, a supernatant group and a blank control group. For each rabbit in fluorometholone group, one cornea was treated with fluorometholone 0.1% eye drops four times a day for 8 weeks, while corneas of rabbits in supernatant group were treated in the same frequency with supernatant fraction centrifuged from fluorometholone 0.1% eye drops. The rabbits in the blank control group were not given any treatment. At the end of the 8 week observation period, the rabbits were euthanized and the eyes immediately enucleated and prepared for inflation testing. The experimental pressure-deformation data was used to derive the stress-strain behavior of each eye using an inverse modeling procedure. Comparisons of mechanical stiffness of corneas were conducted among the three groups to determine the influence of fluorometholone. The results showed that corneal stiffness decreased as the fluorometholone administration time prolonged. Comparisons of tangent modulus indicated average stiffness reductions of 34.2% and 33.5% in the fluorometholone group compared to the supernatant and control groups, respectively, at the end of the observation period. The stiffness-reduction effect of fluorometholone on the cornea should be considered in clinical management, especially when administrating it to biomechanically weakened corneas, such as after refractive surgeries and in cases of keratoconus. © 2013 Published by Elsevier Ltd.

Nerves and neovessels inhibit each other in the cornea.

PubMed

Ferrari, Giulio; Hajrasouliha, Amir R; Sadrai, Zahra; Ueno, Hiroki; Chauhan, Sunil K; Dana, Reza

2013-01-28

To evaluate the regulatory cross-talk of the vascular and neural networks in the cornea. b-FGF micropellets (80 ng) were implanted in the temporal side of the cornea of healthy C57Bl/6 mice. On day 7, blood vessels (hemangiogenesis) and nerves were observed by immunofluorescence staining of corneal flat mounts. The next group of mice underwent either trigeminal stereotactic electrolysis (TSE), or sham operation, to ablate the ophthalmic branch of the trigeminal nerve. Blood vessel growth was detected by immunohistochemistry for PECAM-1 (CD31) following surgery. In another set of mice following TSE or sham operation, corneas were harvested for ELISA (VEGFR3 and pigment epithelium-derived factor [PEDF]) and for quantitative RT-PCR (VEGFR3, PEDF, and CD45). PEDF, VEGFR3, beta-3 tubulin, CD45, CD11b, and F4/80 expression in the cornea were evaluated using immunostaining. No nerves were detected in the areas subject to corneal neovascularization, whereas they persisted in the areas that were neovessel-free. Conversely, 7 days after denervation, significant angiogenesis was detected in the cornea, and this was associated with a significant decrease in VEGFR3 (57.5% reduction, P = 0.001) and PEDF protein expression (64% reduction, P < 0.001). Immunostaining also showed reduced expression of VEGFR3 in the corneal epithelial layer. Finally, an inflammatory cell infiltrate, including macrophages, was observed. Our data suggest that sensory nerves and neovessels inhibit each other in the cornea. When vessel growth is stimulated, nerves disappear and, conversely, denervation induces angiogenesis. This phenomenon, here described in the eye, may have far-reaching implications in understanding angiogenesis.

Validation of an endothelial roll preparation for Descemet Membrane Endothelial Keratoplasty by a cornea bank using "no touch" dissection technique.

PubMed

Marty, Anne-Sophie; Burillon, Carole; Desanlis, Adeline; Damour, Odile; Kocaba, Viridiana; Auxenfans, Céline

2016-06-01

Descemet Membrane Endothelial Keratoplasty (DMEK) selectively replaces the damaged posterior part of the cornea. However, the DMEK technique relies on a manually-performed dissection that is time-consuming, requires training and presents a potential risk of endothelial graft damages leading to surgery postponement when performed by surgeons in the operative room. To validate precut corneal tissue preparation for DMEK provided by a cornea bank in order to supply a quality and security precut endothelial tissue. The protocol was a technology transfer from the Netherlands Institute for Innovative Ocular Surgery (NIIOS) to Lyon Cornea Bank, after formation in NIIOS to the DMEK "no touch" dissection technique. The technique has been validated in selected conditions (materials, microscope) and after a learning curve, cornea bank technicians prepared endothelial tissue for DMEK. Endothelial cells densities (ECD) were evaluated before and after preparation, after storage and transport to the surgery room. Microbiological and histological controls have been done. Twenty corneas were manually dissected; 18 without tears. Nineteen endothelial grafts formed a double roll. The ECD loss after cutting was 3.3 % (n = 19). After transportation 7 days later, we found an ECD loss of 25 % (n = 12). Three days after cutting and transportation, we found 2.1 % of ECD loss (n = 7). Histology found an endothelial cells monolayer lying on Descemet membrane. The mean thickness was 12 ± 2.2 µm (n = 4). No microbial contamination was found (n = 19). Endothelial roll stability has been validated at 3 days in our cornea bank. Cornea bank technicians trained can deliver to surgeons an ECD controlled, safety and ready to use endothelial tissue, for DMEK by "no touch" technique, allowing time saving, quality and security for surgeons.

Effects of new biomimetic regenerating agents on corneal wound healing in an experimental model of post-surgical corneal ulcers.

PubMed

Alcalde, I; Íñigo-Portugués, A; Carreño, N; Riestra, A C; Merayo-Lloves, J M

2015-10-01

The purpose of this study is to assess the effectiveness of the topical application of cacicol regenerating agent (RGTA) in an experimental model of corneal ulcer after photorefractive keratectomy (PRK) in mice. Mice were subjected to PRK surgery with a 2.0mm ablation zone on the central cornea and 45mm of depth on a VISX Star S2 excimer laser. Corneas were treated topically with cacicol drops 1hour and 48hours after injury. Control groups received balanced salt solution (BSS) in the same dosage. Clinical and histopathological events were evaluated at 1, 2, 3 and 7 days after surgery. Sections obtained through the central region of the corneas were used to analyze the histopathological events of injured and healed corneas. αSMA (myofibroblast transformation), E cadherin (assembly of epithelial cells) and neuronal class III β-tubulin (innervation) were performed. Corneas treated topically with cacicol for 7 days showed a greater degree of transparency compared to controls. cacicol treated corneas showed improved epithelial cytoarchitecture. Analysis of αSMA profiles in the stroma showed that cacicol reduced or delayed the presence of myofibroblasts in the stroma compared to BSS (P<0.001). Finally, a putative neuroregenerative effect of cacicol was found in corneas subjected to an experimental PRK lesion. In some cases some interindividual variability could be observed due to the design of the experimental model. This is a limitation to consider, despite the statistical significance of the data. In a model of laser induced surgical lesions in the cornea, topical application of an RGTA (i.e. cacicol) could be involved in avoiding myofibroblast scarring formation and promoting nerve regeneration. Copyright © 2014 Sociedad Española de Oftalmología. Published by Elsevier España, S.L.U. All rights reserved.

EBM regeneration and changes in EBM component mRNA expression in stromal cells after corneal injury

PubMed Central

Santhanam, Abirami; Marino, Gustavo K.; Torricelli, Andre A. M.

2017-01-01

Purpose To investigate the production of the epithelial basement membrane (EBM) component mRNAs at time points before lamina lucida and lamina densa regeneration in anterior stromal cells after corneal injury that would heal with and without fibrosis. Methods Rabbit corneas were removed from 2 to 19 days after −4.5D or −9.0D photorefractive keratectomy (PRK) with the VISX S4 IR laser. Corneas were evaluated with transmission electron microscopy (TEM) for full regeneration of the lamina lucida and the lamina densa. Laser capture microdissection (LCM) based quantitative real-time (RT)–PCR was used to quantitate the expression of mRNAs for laminin α-3 (LAMA3), perlecan, nidogen-1, and nidogen-2 in the anterior stroma. Results After −4.5D PRK, EBM was found to be fully regenerated at 8 to 10 days after surgery. At 4 days after PRK, the nidogen-2 and LAMA3 mRNAs levels were detected at statistically significantly lower levels in the anterior stroma of the −9.0D PRK corneas (where the EBM would not fully regenerate) compared to the −4.5D PRK corneas (where the EBM was destined to fully regenerate). At 7 days after PRK, nidogen-2 and LAMA3 mRNAs continued to be statistically significantly lower in the anterior stroma of the −9.0D PRK corneas compared to their expression in the anterior stroma of the −4.5D PRK corneas. Conclusions Key EBM components LAMA3 and nidogen-2 mRNAs are expressed at higher levels in the anterior stroma during EBM regeneration in the −4.5D PRK corneas where the EBM is destined to fully regenerate and no haze developed compared to the −9.0D PRK corneas where the EBM will not fully regenerate and myofibroblast-related stromal fibrosis (haze) will develop. PMID:28275314

Oxygen transport through soft contact lens and cornea: Lens characterization and metabolic modeling

NASA Astrophysics Data System (ADS)

Chhabra, Mahendra

The human cornea requires oxygen to sustain metabolic processes critical for its normal functioning. Any restriction to corneal oxygen supply from the external environment (e.g., by wearing a low oxygen-permeability contact lens) can lead to hypoxia, which may cause corneal edema (swelling), limbal hyperemia, neovascularization, and corneal acidosis. The need for adequate oxygen to the cornea is a major driving force for research and development of hypertransmissible soft contact lenses (SCLs). Currently, there is no standard technique for measuring oxygen permeability (Dk) of hypertransmissible silicone-hydrogel SCLs. In this work, an electrochemistry-based polarographic apparatus was designed, built, and operated to measure oxygen permeability in hypertransmissible SCLs. Unlike conventional methods where a range of lens thickness is needed for determining oxygen permeabilities of SCLs, this apparatus requires only a single lens thickness. The single-lens permeameter provides a reliable, efficient, and economic tool for measuring oxygen permeabilities of commercial hypertransmissible SCLs. The single-lens permeameter measures not only the product Dk, but, following modification, it measures separately diffusivity, D, and solubility, k, of oxygen in hypertransmissible SCLs. These properties are critical for designing better lens materials that ensure sufficient oxygen supply to the cornea. Metabolism of oxygen in the cornea is influenced by contact-lens-induced hypoxia, diseases such as diabetes, surgery, and drug treatment, Thus, estimation of the in-vivo corneal oxygen consumption rate is essential for gauging adequate oxygen supply to the cornea. Therefore, we have developed an unsteady-state reactive-diffusion model for the cornea-contact-lens system to determine in-vivo human corneal oxygen-consumption rate. Finally, a metabolic model was developed to determine the relation between contact-lens oxygen transmissibility (Dk/L) and corneal oxygen deficiency. A new parameter "Oxygen Deficiency Factor" (ODF) is defined to quantify oxygen deficiency in local regions of the cornea. We use this concept to determine the minimum required contact-lens oxygen transmissibility, Dk/L = 150 Barrer/cm, to avoid hypoxia-induced corneal physiologic complications.

[The value of methods for morphofunctional analysis of cornea in cataract surgery].

PubMed

Borodina, N B; Kobzova, M V; Musaeva, G M

2011-01-01

The characteristics of morphofunctional status of cornea after extracapsular cataract extraction and phakoemulsification with IOL implantation (30 and 58 operations respectively) were analyzed in detail using up-to-date diagnostic techniques. The results of examination using developed algorithm including study of light transmission, refraction and protective function of cornea show advantage of microinvasive ultrasound technique of cataract surgery in terms of minimal impact on corneal structure, optical and biomechanical characteristics.

Pediatric genetic disease of the cornea

PubMed Central

Fecarotta, Christopher M.; Huang, Wendy W.

2014-01-01

Our objective is to evaluate the literature regarding selected genetic diseases of the cornea, including megalocornea, keratoglobus, keratoconus, cystinosis, the mucopolysaccharidoses, sclerocornea, Peters' anomaly, familial dysautonomia, and various corneal dystrophies. The transparency of the cornea is a consequence of uniformity in both size and spacing of the collagen lamellae. The cornea's clarity depends on a delicate biochemical and structural balance; consequently, genetic disorders that disrupt either its metabolic or anatomic function can cause opacity and vision loss. Many childhood corneal diseases have a genetic etiology and are associated with known syndromes. Each disorder has unique associated set of possible complications. Prognosis often depends on the extent of opacity and disorganization of the anterior segment. Corneal transplantation has been performed for these disorders with variable success. PMID:27625877

Surveillance of Vittaforma corneae in hot springs by a small-volume procedure.

PubMed

Chen, Jung-Sheng; Hsu, Tsui-Kang; Hsu, Bing-Mu; Huang, Tung-Yi; Huang, Yu-Li; Shaio, Men-Fang; Ji, Dar-Der

2017-07-01

Vittaforma corneae is an obligate intracellular fungus and can cause human ocular microsporidiosis. Although accumulating reports of V. corneae causing keratoconjunctivitis in both healthy and immunocompromised persons have been published, little is known about the organism's occurrence in aquatic environments. Limitations in detection sensitivity have meant a large sampling volume is required to detect the pathogen up to now, which is problematic. A recent study in Taiwan has shown that some individuals suffering from microsporidial keratitis (MK) were infected after exposure to the pathogen at a hot spring. As a consequence of this, a survey and analysis of environmental V. corneae present in hot springs became an urgent need. In this study, sixty water samples from six hot spring recreation areas around Taiwan were analyzed. One liter of water from each sample site was filtered to harvest the fungi. The positive samples were detected using a modified nested PCR approach followed by sequencing using specific SSU rRNA gene primer pairs for V. corneae. In total fifteen V. corneae-like isolates were identified (25.0% of sites). Among them, six isolates, which were collected from recreational areas B, C and D, were highly similar to known V. corneae keratitis strains from Taiwan and other countries. Furthermore, five isolates, which were collected from recreation areas A, C, E and F, were very similar to Vittaforma-like diarrhea strains isolated in Portugal. Cold spring water tubs and public foot bath pools had the highest detection rate (50%), suggesting that hot springs might be contaminated via untreated water sources. Comparing the detection rate across different regions of Taiwan, Taitung, which is in the east of the island, gave the highest positive rate (37.5%). Statistical analysis showed that outdoor/soil exposure and a high heterotrophic plate count (HPC) were risk factors for the occurrence of V. corneae. Our findings provide empirical evidence supporting the need for proper control and regulations at hot spring recreational waters in order to avoid health risks from this pathogen. Finally, we have developed a small volume procedure for detecting V. corneae in water samples and this has proved to be very useful. Copyright © 2017 Elsevier Ltd. All rights reserved.

Rocking media over ex vivo corneas improves this model and allows the study of the effect of proinflammatory cytokines on wound healing.

PubMed

Deshpande, Pallavi; Ortega, Ílida; Sefat, Farshid; Sangwan, Virender S; Green, Nicola; Claeyssens, Frederik; MacNeil, Sheila

2015-02-05

The aim of this work was to develop an in vitro cornea model to study the effect of proinflammatory cytokines on wound healing. Initial studies investigated how to maintain the ex vivo models for up to 4 weeks without loss of epithelium. To study the effect of cytokines, corneas were cultured with the interleukins IL-17A, IL-22, or a combination of IL-17A and IL-22, or lipopolysaccharide (LPS). The effect of IL-17A on wound healing was then examined. With static culture conditions, organ cultures deteriorated within 2 weeks. With gentle rocking of media over the corneas and carbon dioxide perfusion, the ex vivo models survived for up to 4 weeks without loss of epithelium. The cytokine that caused the most damage to the cornea was IL-17A. Under static conditions, wound healing of the central corneal epithelium occurred within 9 days, but only a single-layered epithelium formed whether the cornea was exposed to IL-17A or not. With rocking of media gently over the corneas, a multilayered epithelium was achieved 9 days after wounding. In the presence of IL-17A, however, there was no wound healing evident. Characterization of the cells showed that wherever epithelium was present, both differentiated cells and highly proliferative cells were present. We propose that introducing rocking to extend the effective working life of this model and the introduction of IL-17A to this model to induce aspects of inflammation extend its usefulness to study the effects of agents that influence corneal regeneration under normal and inflamed conditions. Copyright 2015 The Association for Research in Vision and Ophthalmology, Inc.

Elastic microfibril distribution in the cornea: Differences between normal and keratoconic stroma.

PubMed

White, Tomas L; Lewis, Philip N; Young, Robert D; Kitazawa, Koji; Inatomi, Tsutomu; Kinoshita, Shigeru; Meek, Keith M

2017-06-01

The optical and biomechanical properties of the cornea are largely governed by the collagen-rich stroma, a layer that represents approximately 90% of the total thickness. Within the stroma, the specific arrangement of superimposed lamellae provides the tissue with tensile strength, whilst the spatial arrangement of individual collagen fibrils within the lamellae confers transparency. In keratoconus, this precise stromal arrangement is lost, resulting in ectasia and visual impairment. In the normal cornea, we previously characterised the three-dimensional arrangement of an elastic fiber network spanning the posterior stroma from limbus-to-limbus. In the peripheral cornea/limbus there are elastin-containing sheets or broad fibers, most of which become microfibril bundles (MBs) with little or no elastin component when reaching the central cornea. The purpose of the current study was to compare this network with the elastic fiber distribution in post-surgical keratoconic corneal buttons, using serial block face scanning electron microscopy and transmission electron microscopy. We have demonstrated that the MB distribution is very different in keratoconus. MBs are absent from a region of stroma anterior to Descemet's membrane, an area that is densely populated in normal cornea, whilst being concentrated below the epithelium, an area in which they are absent in normal cornea. We contend that these latter microfibrils are produced as a biomechanical response to provide additional strength to the anterior stroma in order to prevent tissue rupture at the apex of the cone. A lack of MBs anterior to Descemet's membrane in keratoconus would alter the biomechanical properties of the tissue, potentially contributing to the pathogenesis of the disease. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Effect of glaucoma tube shunt parameters on cornea endothelial cells in patients with Ahmed valve implants.

PubMed

Koo, Euna B; Hou, Jing; Han, Ying; Keenan, Jeremy D; Stamper, Robert L; Jeng, Bennie H

2015-01-01

The aim of this study was to assess the effect of various tube parameters on corneal endothelial cell density (ECD) after insertion of Ahmed valves. Thirty-nine eyes of 33 patients with previous superotemporal (ST) Ahmed valve implantation and 20 eyes of 13 participants with previous uncomplicated phacoemulsification and intraocular lens implantation but no history of glaucoma surgery were evaluated. Various tube parameters were measured with anterior segment optical coherence tomography. ST, central, and inferonasal (IN) ECD and pachymetry were measured. Endothelial cell loss and corneal thickness in the ST cornea was compared with those in the IN cornea. The mean age of the operated patients was 58 ± 22 years, and the mean time since glaucoma surgery was 2.5 ± 2.6 years. Thirty-two of the 39 study eyes were pseudophakic. The ECD was significantly lower in the ST endothelium than in the IN endothelium in eyes with glaucoma tube surgery (P < 0.001), although this relative reduction in ST ECD was not greater than that seen in pseudophakic control eyes (P = 0.16). In univariate analysis, tube angle relative to the cornea and distance from the tip of the tube to the cornea were significant risk factors for decreased ST endothelial cell loss when assessed relative to the IN ECD (P = 0.01 and P = 0.02, respectively). In multivariate analysis, only the distance of the tube tip to the cornea remained significantly associated with ST endothelial cell loss. Although this was a retrospective study with inherent limitations, tubes that are closer to the cornea seem to lead to increased loss of adjacent endothelial cells.

Characterization of Ocular Iontophoretic Drug Transport of Ionic and Non-ionic Compounds in Isolated Rabbit Cornea and Conjunctiva.

PubMed

Sekijima, Hidehisa; Ehara, Junya; Hanabata, Yusuke; Suzuki, Takumi; Kimura, Soichiro; Lee, Vincent H L; Morimoto, Yasunori; Ueda, Hideo

2016-06-01

Ocular iontophoresis (IP) in isolated rabbit cornea and conjunctiva was examined in terms of transport enhancement, tissue viability and integrity using electrophysiological parameters by the Ussing-type chamber technique. Lidocaine hydrochloride (LC, a cationic compound), sodium benzoate (BA, anionic compound), and fluorescein isothiocyanate labeled dextran (molecular weight 4400 Da, FD-4, hydrophilic large compound) were used as model permeants. Direct electric current was applied at 0.5-5.0 mA/cm(2) for the cornea and 0.5-20 mA/cm(2) for the conjunctiva for 30 min. LC and BA fluxes across the cornea and conjunctiva were significantly increased by the application of electric current up to 2.3- and 2.5-fold and 4.0- and 3.4-fold, respectively, and returned to their baseline level on stopping the current. Furthermore, a much higher increase by IP application was obtained for the FD-4 transport. The increased FD-4 flux in the conjunctiva returned to baseline on stopping the current, whereas the flux in the cornea was sustained at a higher level after stopping the current. The transepithelial electric resistance of the cornea and conjunctiva was lowered by electric current application but fully recovered after stopping the current up to 2.0 mA/cm(2) for the cornea and 10 mA/cm(2) for the conjunctiva, suggesting that the corneal and conjunctival viability and integrity are maintained even after application of these current densities. These results indicate that ocular IP may be a useful non-invasive technique to achieve drug delivery of hydrophilic large molecules into the eyes.

Mincle in the innate immune response of mice fungal keratitis.

PubMed

Yu, Guo-Rong; Lin, Jing; Zhang, Jie; Che, Cheng-Ye; Peng, Xu-Dong; Li, Cui; Hu, Li-Ting; Zhu, Guo-Qiang; He, Kun; Zhao, Gui-Qiu

2018-01-01

To investigate how macrophage inducible C-type lectin (Mincle) influences inflammation in mice fungal keratitis induced by Aspergillus fumigatus ( A. fumigatus ). C57BL/6 mice were infected with A. fumigatus after pretreated with Mincle agonist TDB or Mincle neutralizing antibody (MincleAb), taking DMSO or IgG as control group respectively. The cornea lesions were monitored with slit-lamp microscope and evaluated by clinical score. Mincle expression was assessed using reverse transcription-ploymerase chain reaction (RT-PCR) and immunostaining. The expression of cytokines (IL-1β, TNF-α and IL-6) chemokines (CXCL-1 and MIP-2) was determined by RT-PCR and ELISA. Neutrophil infiltration was observed by immunostaining. The levels of nitric oxide (NO) generated by corneas were tested by Griess reaction. Mincle mRNA and protein levels were higher in infected corneas than normal corneas of C57BL/6 mice, saving clinical scores revealed differences. When pretreated with Mincle agonist TDB, the mRNA and protein levels of IL-1β, TNF-α and IL-6 in infected corneas were significantly increased compared with the control group ( P <0.01). Results of the counterpart in corneas pretreated with Mincle neutralizing antibody was decreased consistently ( P <0.01). Expression of CXCL1 and MIP-2 mRNA levels were up-regulated in TDB group and down-regulated in MincleAb group ( P <0.01), coincide with neutrophil aggregation degree in corneas showed by immunostaining. As for the concentration of NO, it was promoted in TDB group compared with DMSO control group, and decreased in MincleAb group compared with IgG control group. Mincle plays a dual role in mice fungal keratitis. It participates in the innate immune system by enhancing inflammation. What's more, Mincle can mediate cytotoxic effects by regulating the formation of NO.

Reliability of Pentacam HR Thickness Maps of the Entire Cornea in Normal, Post-Laser In Situ Keratomileusis, and Keratoconus Eyes.

PubMed

Xu, Zhe; Peng, Mei; Jiang, Jun; Yang, Chun; Zhu, Weigen; Lu, Fan; Shen, Meixiao

2016-02-01

To measure the repeatability and reproducibility of Pentacam HR system thickness maps for the entire cornea in normal, post-laser in situ keratomileusis (post-LASIK), and keratoconus (KC) eyes. Reliability study. Sixty normal subjects (60 eyes), 30 post-LASIK subjects (60 eyes), and 14 KC patients (27 eyes) were imaged with the Pentacam HR system by 2 well-trained operators. For pachymetry the cornea was divided into 4 zones: a central zone (2-mm diameter) and concentric pericentral zone (2-5 mm), transitional zone (5-7 mm), and peripheral zone (7-10 mm). The 3 concentric zones were subdivided into 8 sectors. Intraobserver repeatability and interobserver reproducibility of entire corneal thickness maps were tested by the repeatability and reproducibility coefficients, intraclass correlation coefficients, coefficient of variation, and 95% limits of agreement. From central to peripheral zones, the precision of corneal thickness measurements became gradually smaller. Central zone repeatability and reproducibility were the best in the normal, post-LASIK, and KC groups. The peripheral superior sectors showed poorer repeatability and reproducibility for all subjects. The intraobserver repeatability and interobserver reproducibility for all zones were ≤19.3 μm, ≤22.1 μm, and ≤20.7 μm, in the normal, post-LASIK, and KC groups, respectively. The intraobserver and interobserver coefficients of variation for all zones were ≤1.3%, ≤1.6%, and ≤1.6% for all 3 groups. Pentacam HR system pachymetry of the entire cornea provided good precision in normal, post-LASIK, and KC corneas. Thickness measurements in the peripheral cornea should be interpreted with caution in abnormal corneas after surgery or with diseases. Copyright © 2016 Elsevier Inc. All rights reserved.

Nerves and Neovessels Inhibit Each Other in the Cornea

PubMed Central

Ferrari, Giulio; Hajrasouliha, Amir R.; Sadrai, Zahra; Ueno, Hiroki; Chauhan, Sunil K.; Dana, Reza

2013-01-01

Purpose. To evaluate the regulatory cross-talk of the vascular and neural networks in the cornea. Methods. b-FGF micropellets (80 ng) were implanted in the temporal side of the cornea of healthy C57Bl/6 mice. On day 7, blood vessels (hemangiogenesis) and nerves were observed by immunofluorescence staining of corneal flat mounts. The next group of mice underwent either trigeminal stereotactic electrolysis (TSE), or sham operation, to ablate the ophthalmic branch of the trigeminal nerve. Blood vessel growth was detected by immunohistochemistry for PECAM-1 (CD31) following surgery. In another set of mice following TSE or sham operation, corneas were harvested for ELISA (VEGFR3 and pigment epithelium-derived factor [PEDF]) and for quantitative RT-PCR (VEGFR3, PEDF, and CD45). PEDF, VEGFR3, beta-3 tubulin, CD45, CD11b, and F4/80 expression in the cornea were evaluated using immunostaining. Results. No nerves were detected in the areas subject to corneal neovascularization, whereas they persisted in the areas that were neovessel-free. Conversely, 7 days after denervation, significant angiogenesis was detected in the cornea, and this was associated with a significant decrease in VEGFR3 (57.5% reduction, P = 0.001) and PEDF protein expression (64% reduction, P < 0.001). Immunostaining also showed reduced expression of VEGFR3 in the corneal epithelial layer. Finally, an inflammatory cell infiltrate, including macrophages, was observed. Conclusion. Our data suggest that sensory nerves and neovessels inhibit each other in the cornea. When vessel growth is stimulated, nerves disappear and, conversely, denervation induces angiogenesis. This phenomenon, here described in the eye, may have far-reaching implications in understanding angiogenesis. PMID:23307967

Non-contact full-field optical coherence tomography: a novel tool for in vivo imaging of the human cornea (Conference Presentation)

NASA Astrophysics Data System (ADS)

Mazlin, Viacheslav; Dalimier, Eugénie; Grieve, Katharine F.; Irsch, Kristina; Sahel, José-Alain; Fink, Mathias; Boccara, A. Claude

2017-02-01

According to the World Health Organization (WHO), corneal diseases alongside with cataract and retinal diseases are major causes of blindness worldwide. For the 95.5% of corneal blindness cases, prevention or rehabilitation could have been possible without negative consequences for vision, provided that disease is diagnosed early. However, diagnostics at the early stage requires cellular-level resolution, which is not achieved with routinely used Slit-lamp and OCT instruments. Confocal microscopy allows examination of the cornea at a resolution approaching histological detail, however requires contact with a patient's eye. The recently developed full-field OCT technique, in which 2D en face tangential optical slices are directly recorded on a camera, was successfully applied for ex vivo eye imaging. However, in vivo human eye imaging has not been demonstrated yet. Here we present a novel non-contact full-field OCT system, which is capable of imaging in air and, therefore, shows potential for in vivo cornea imaging in patients. The first cellular-level resolution ex vivo images of cornea, obtained in a completely non-contact way, were demonstrated. We were able to scan through the entire cornea (400 µm) and resolve epithelium, Bowman's layer, stroma and endothelium. FFOCT images of the human cornea in vivo were obtained for the first time. The epithelium structures and stromal keratocyte cells were distinguishable. Both ex vivo and in vivo images were acquired with a large (1.26 mm x 1.26 mm) field of view. Cellular details in obtained images make this device a promising candidate for realization of high-resolution in vivo cornea imaging.

Identification and characterization of a Na+-dependent neutral amino acid transporter, ASCT1, in rabbit corneal epithelial cell culture and rabbit cornea.

PubMed

Katragadda, Suresh; Talluri, Ravi Sankar; Pal, Dhananjay; Mitra, Ashim K

2005-11-01

The aim of this study was to investigate the presence of a Na+-dependent neutral amino acid transporter, ASCT1, in rabbit primary corneal epithelial cell culture and rabbit cornea. Uptake studies were carried out on rabbit primary corneal epithelial culture (rPCEC) cells using 12-well plates. Transport studies were conducted with isolated rabbit corneas at 34 degrees C. Uptake and transport of L-alanine was determined at various concentrations. Inhibition studies were conducted in presence of various L- and D-amino acids, metabolic inhibitors like ouabain and sodium azide, and in the absence of sodium to delineate the functional characteristics of L-alanine uptake and transport. Reverse transcription-polymerase chain reaction (RT-PCR) was performed on total RNA harvested from rabbit cornea and rPCEC cells for identification of ASCT1. Uptake of L-Ala was found to be saturable with a Km of 0.71 mM and a Vmax value of 0.84 micromoles min(-1) mg(-1) protein. Uptake was independent of pH and energy but depends on sodium. It was inhibited by serine, threonine, cysteine, and glutamine but did not respond to BCH (2-aminobicyclo [2,2,1] heptane-2-carboxylic acid) and MeAIB (alpha -methylaminoisobutyric acid). Transport of L-Ala across rabbit cornea was also saturable (Km 6.52 mM and Vmax 1.09 x 10(-2) micromoles min(-1) cm(-2)), energy independent, and subject to similar competitive inhibition. Presence of ASCT1 on rPCEC and on rabbit cornea was identified by RT-PCR. L-Alanine, the chosen model substrate, was actively transported by Na+-dependent, neutral amino acid exchanger ASCT1, which was identified and functionally characterized on rPCEC cells and rabbit cornea.

Public Health Implications of the 1540 nm Laser on the Cornea

DTIC Science & Technology

2007-05-30

film layer. The epithelium has numerous nerve endings that make the cornea extremely sensitive to pain when an injury occurs. This pain reflex helps...used in research by using techniques and procedures that reduce distress and pain The Three R’s have lead researchers to look for alternative models...cause severe pain and render a person incapable of working. Injury to the cornea may cause temporary to permanent vision impairment depending on how

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sobol', E N; Vorobieva, N N; Omel'chenko, A I

A new approach is proposed for correcting the eye refraction by controlled variation of the mechanical properties of the sclera and cornea upon nondestructive laser heating. Experimental ex vivo studies of rabbit and pig eyes show that laser-induced local denaturation of the sclera changes the refraction of the cornea by 3 diopters on the average, and the subsequent nondestructive irradiation of the cornea increases its plasticity, which leads to a further increase in its radius of curvature and a decrease in refraction down to 7 diopters.

Transverse depth-dependent changes in corneal collagen lamellar orientation and distribution

PubMed Central

Abass, Ahmed; Hayes, Sally; White, Nick; Sorensen, Thomas; Meek, Keith M.

2015-01-01

It is thought that corneal surface topography may be stabilized by the angular orientation of out-of plane lamellae that insert into the anterior limiting membrane. In this study, micro-focus X-ray scattering data were used to obtain quantitative information about lamellar inclination (with respect to the corneal surface) and the X-ray scatter intensity throughout the depth of the cornea from the centre to the temporal limbus. The average collagen inclination remained predominantly parallel to the tissue surface at all depths. However, in the central cornea, the spread of inclination angles was greatest in the anterior-most stroma (reflecting the increased lamellar interweaving in this region), and decreased with tissue depth; in the peripheral cornea inclination angles showed less variation throughout the tissue thickness. Inclination angles in the deeper stroma were generally higher in the peripheral cornea, suggesting the presence of more interweaving in the posterior stroma away from the central cornea. An increase in collagen X-ray scatter was identified in a region extending from the sclera anteriorly until about 2 mm from the corneal centre. This could arise from the presence of larger diameter fibrils, probably of scleral origin, which are known to exist in this region. Incorporation of this quantitative information into finite-element models will further improve the accuracy with which they can predict the biomechanical response of the cornea to pathology and refractive procedures. PMID:25631562

b-FGF induces corneal blood and lymphatic vessel growth in a spatially distinct pattern.

PubMed

Hajrasouliha, Amir R; Sadrai, Zahra; Chauhan, Sunil K; Dana, Reza

2012-07-01

To study the spatial variances in ligand expression and angiogenic effect in response to the inflammatory response induced by basic fibroblast growth factor (b-FGF). b-FGF micropellets (80 ng) were implanted in the temporal side of the cornea of Balb/c mice. On days 1, 3, and 7, blood (heme-) and lymphangiogenesis were observed by immunofluorescence staining of corneal flat mounts with LYVE-1 and CD31 to identify lymphatic and blood vessels, respectively. A second group of corneas were harvested for quantitative real-time polymerase chain reaction. Each cornea was divided into 2 different areas: (1) pre-pellet area and (2) opposite-pellet area. Expression of vascular endothelial growth factor (VEGF) ligands was evaluated using real-time polymerase chain reaction in each respective zone. Blood vessels grew into the cornea from the pre-pellet area, whereas corneal lymphatic vessels grew from the opposite-pellet area toward the center of the cornea. VEGF-A was upregulated in the pre-pellet, whereas VEGF-D expression was mostly observed in the opposite-pellet area. VEGF-C level increased simultaneously in both areas. A single inducing factor, that is, b-FGF, may simultaneously provoke hemangiogenesis and lymphangiogenesis in different locations of the cornea through differential expression of VEGF ligands. This distinctive spatial pattern should be considered while evaluating the corneal predilection for inflammation beyond that which is directly visible by slit lamp examination.

Patient-specific simulation of the intrastromal ring segment implantation in corneas with keratoconus.

PubMed

Lago, M A; Rupérez, M J; Monserrat, C; Martínez-Martínez, F; Martínez-Sanchis, S; Larra, E; Díez-Ajenjo, M A; Peris-Martínez, C

2015-11-01

The purpose of this study was the simulation of the implantation of intrastromal corneal-ring segments for patients with keratoconus. The aim of the study was the prediction of the corneal curvature recovery after this intervention. Seven patients with keratoconus diagnosed and treated by implantation of intrastromal corneal-ring segments were enrolled in the study. The 3D geometry of the cornea of each patient was obtained from its specific topography and a hyperelastic model was assumed to characterize its mechanical behavior. To simulate the intervention, the intrastromal corneal-ring segments were modeled and placed at the same location at which they were placed in the surgery. The finite element method was then used to obtain a simulation of the deformation of the cornea after the ring segment insertion. Finally, the predicted curvature was compared with the real curvature after the intervention. The simulation of the ring segment insertion was validated comparing the curvature change with the data after the surgery. Results showed a flattening of the cornea which was in consonance with the real improvement of the corneal curvature. The mean difference obtained was of 0.74 mm using properties of healthy corneas. For the first time, a patient-specific model of the cornea has been used to predict the outcomes of the surgery after the intrastromal corneal-ring segments implantation in real patients. Copyright © 2015 Elsevier Ltd. All rights reserved.

Ex vivo rabbit cornea diffusion studies with a soluble insert of moxifloxacin.

PubMed

Sebastián-Morelló, María; Calatayud-Pascual, María Aracely; Rodilla, Vicent; Balaguer-Fernández, Cristina; López-Castellano, Alicia

2018-02-01

The objective of this research was to develop and evaluate an ocular insert for the controlled drug delivery of moxifloxacin which could perhaps be used in the treatment of corneal keratitis or even bacterial endophthalmitis. We have evaluated the ex vivo ocular diffusion of moxifloxacin through rabbit cornea, both fresh and preserved under different conditions. Histological studies were also carried out. Subsequently, drug matrix inserts were prepared using bioadhesive polymers. The inserts were evaluated for their physicochemical parameters. Ophthalmic ex vivo permeation of moxifloxacin was carried out with the most promising insert. The formulate insert was thin and provided higher ocular diffusion than commercial formulations. Ocular diffusion studies revealed significant differences between fresh and frozen corneas. Histological examinations also showed differences in the thickness of stroma between fresh and frozen corneas. The ophthalmic insert we have developed allows a larger quantity of moxifloxacin to permeate through the cornea than existing commercial formulations of the drug. Ocular delivery of moxifloxacin with this insert could be a new approach for the treatment of eye diseases.

Lymphatic vessels correlate closely with inflammation index in alkali burned cornea.

PubMed

Yan, Hao; Qi, Chaoxiu; Ling, Shiqi; Li, Weihua; Liang, Linyi

2010-08-01

To study the relationship between corneal lymphangiogenesis and inflammation in alkali burned corneas. Rat corneal lymphatic and blood vessels were labeled and distinguished by whole mount immunofluorescence and 5'-nase-alkaline phosphatase (5'-NA-ALP) double enzyme-histochemistry. Then, lymphatic vessel areas (LVA) and lymphatic vessel counting (LVC) were examined. Corneal inflammation was evaluated by inflammation index (IF) grading, histopathology, electron microscope, and polymorphonuclear leukocyte (PMN) infiltration. The relationship between LVC, LVA, IF, and PMN was examined, respectively. In addition, corneal lymphatic vessels of eleven human alkali burned corneas were examined by lymphatic vessel endothelial receptor (LYVE-1) immunohistochemistry. Corneal lymphangiogenesis occurred on Day 3, reached the peak at the end of two weeks, and disappeared five weeks after alkaline burns. Both LVA and LVC were strongly and positively correlated with IF after corneal alkaline burns. However, the relationship between LVC and PMN, between LVA and PMN were significant but converse. Among eleven human alkali burned corneas, corneal lymphangiogenesis was present in three corneas. Corneal lymphagiogenesis develops after alkaline burns and correlates closely with corneal inflammation.

Corneal critical barrier against the penetration of dexamethasone and lomefloxacin hydrochloride: evaluation by the activation energy for drug partition and diffusion in cornea.

PubMed

Yasueda, Shin-ichi; Higashiyama, Masayo; Yamaguchi, Masazumi; Isowaki, Akiharu; Ohtori, Akira

2007-08-01

The cornea is a solid barrier against drug permeation. We searched the critical barrier of corneal drug permeation using a hydrophobic drug, dexamethasone (DM), and a hydrophilic drug, lomefloxacin hydrochloride (LFLX). The activation energies for permeability of DM and LFLX across the intact cornea were 88.0 and 42.1 kJ/mol, respectively. Their activation energies for permeability across the cornea without epithelium decreased to 33.1 and 16.6 kJ/mol, respectively. The results show that epithelium is the critical barrier on the cornea against the permeation of a hydrophobic drug of DM as well as a hydrophilic drug of LFLX. The activation energy of partition for DM (66.8 kJ/mol) was approximately 3-fold larger than that of diffusion (21.2 kJ/mol). The results indicate that the partition for the hydrophobic drug of DM to the corneal epithelium is the primary barrier. Thermodynamic evaluation of activation energy for the drug permeation parameters is a good approach to investigate the mechanism of drug permeability.

Recovery From Amiodarone-Induced Cornea Verticillata by Application of Topical Heparin.

PubMed

Frings, Andreas; Schargus, Marc

2017-11-01

To report a case of amiodarone-induced vortex keratopathy-associated anatomical findings and subjective visual perception before and after treatment with topical heparin eye drops. Case report. A 76-year-old man complained of halos in his vision in both his eyes due to prominent bilateral cornea verticillata. For treatment of cornea verticillata, we prescribed unpreserved eye drops of a sterile, phosphate-free solution of 0.1% sodium hyaluronate with 1300 IU/mL heparin sodium 3 times daily to the left eye, whereas the other side served as the control. The area of corneal deposits was measured by 2 examiners before and at the 1- and 3-month examination. At last follow-up, cornea verticillata had been reduced from 6 to 2 mm in area by approximately 66% from grade-III to grade-II amiodarone keratopathy. In patients using amiodarone, clearing of cornea verticillata may be achieved by topical use of unpreserved eye drops of a sterile, phosphate-free solution of 0.1% sodium hyaluronate with 1300 IU/mL heparin sodium.

Cornea Transplant

MedlinePlus

... transplants may be used that remove only certain layers of cornea tissue, or only tissue affected by ... procedure removes diseased tissue from the back corneal layers, including the endothelium, along with the Descemet membrane, ...

Corneal imaging by second and third harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Brocas, Arnaud; Jay, Louis; Mottay, Eric; Brunette, Isabelle; Ozaki, Tsuneyuki

2008-02-01

Advanced imaging methods are essential tools for improved outcome of refractive surgery. Second harmonic generation (SHG) and third harmonic generation (THG) microscopy are noninvasive high-resolution imaging methods, which can discriminate the different layers of the cornea, thus having strong impact on the outcome of laser surgery. In this work, we use an Ytterbium femtosecond laser as the laser source, the longer wavelength of which reduces scattering, and allows simultaneous SHG and THG imaging. We present SHG and THG images and profiles of pig corneas that clearly show the anterior surface of the cornea, the entry in the stroma and its end, and the posterior surface of the cornea. These observations allow localizing the epithelium, the stroma and the endothelium. Other experiments give information about the structure and cytology of the corneal layers.

Apparent respiration rate of the human corneal epithelium with tetracaine HCl and benoxinate HCl.

PubMed

Bentley, C R; Larke, J R

1983-12-01

Local anesthetics may have a cytotoxic effect which causes a depression in the apparent epithelial oxygen uptake rate (AEOR) of the cornea. We measured the AEOR of human corneas in vivo before and after applying 1% tetracaine (amethocaine) HCl and 0.4% benoxinate HCl. These drugs had no effect on AEOR. In human corneas that had been subjected to a period of hypoxia, AEOR was slightly higher after administration of benoxinate, a result in the opposite direction to that expected on the grounds of toxicity. The increase was not statistically significant. We conclude that clinical doses of tetracaine HCl and benoxinate HCl normally have a minimal cytotoxic effect, and that this is similarly true when benoxinate is applied to the cornea after contact lens wear.

A comparison of glycosaminoglycan distributions, keratan sulphate sulphation patterns and collagen fibril architecture from central to peripheral regions of the bovine cornea

PubMed Central

Ho, Leona T.Y.; Harris, Anthony M.; Tanioka, Hidetoshi; Yagi, Naoto; Kinoshita, Shigeru; Caterson, Bruce; Quantock, Andrew J.; Young, Robert D.; Meek, Keith M.

2014-01-01

This study investigated changes in collagen fibril architecture and the sulphation status of keratan sulphate (KS) glycosaminoglycan (GAG) epitopes from central to peripheral corneal regions. Freshly excised adult bovine corneal tissue was examined as a function of radial position from the centre of the cornea outwards. Corneal thickness, tissue hydration, hydroxyproline content, and the total amount of sulphated GAG were all measured. High and low-sulphated epitopes of keratan sulphate were studied by immunohistochemistry and quantified by ELISA. Chondroitin sulphate (CS) and dermatan sulphate (DS) distributions were observed by immunohistochemistry following specific enzyme digestions. Electron microscopy and X-ray fibre diffraction were used to ascertain collagen fibril architecture. The bovine cornea was 1021 ± 5.42 μm thick at its outer periphery, defined as 9–12 mm from the corneal centre, compared to 844 ± 8.10 μm at the centre. The outer periphery of the cornea was marginally, but not significantly, more hydrated than the centre (H = 4.3 vs. H = 3.7), and was more abundant in hydroxyproline (0.12 vs. 0.06 mg/mg dry weight of cornea). DMMB assays indicated no change in the total amount of sulphated GAG across the cornea. Immunohistochemistry revealed the presence of both high- and low-sulphated epitopes of KS, as well as DS, throughout the cornea, and CS only in the peripheral cornea before the limbus. Quantification by ELISA, disclosed that although both high- and low-sulphated KS remained constant throughout stromal depth at different radial positions, high-sulphated epitopes remained constant from the corneal centre to outer-periphery, whereas low-sulphated epitopes increased significantly. Both small angle X-ray diffraction and TEM analysis revealed that collagen fibril diameter remained relatively constant until the outer periphery was reached, after which fibrils became more widely spaced (from small angle x-ray diffraction analysis) and of larger diameter as they approached the sclera. Depth-profiled synchrotron microbeam analyses showed that, at different radial positions from the corneal centre outwards, fibril diameter was greater superficially than in deeper stromal regions. The interfibrillar spacing was also higher at mid-depth in the stroma than it was in anterior and posterior stromal regions. Collagen fibrils in the bovine cornea exhibited a fairly consistent spacing and diameter from the corneal centre to the 12 mm radial position, after which a significant increase was seen. While the constancy of the overall sulphation levels of proteoglycans in the cornea may correlate with the fibrillar architecture, there was no correlation between the latter and the distribution of low-sulphated KS. PMID:25019467

Algorithms for High-Speed Noninvasive Eye-Tracking System

NASA Technical Reports Server (NTRS)

Talukder, Ashit; Morookian, John-Michael; Lambert, James

2010-01-01

Two image-data-processing algorithms are essential to the successful operation of a system of electronic hardware and software that noninvasively tracks the direction of a person s gaze in real time. The system was described in High-Speed Noninvasive Eye-Tracking System (NPO-30700) NASA Tech Briefs, Vol. 31, No. 8 (August 2007), page 51. To recapitulate from the cited article: Like prior commercial noninvasive eyetracking systems, this system is based on (1) illumination of an eye by a low-power infrared light-emitting diode (LED); (2) acquisition of video images of the pupil, iris, and cornea in the reflected infrared light; (3) digitization of the images; and (4) processing the digital image data to determine the direction of gaze from the centroids of the pupil and cornea in the images. Most of the prior commercial noninvasive eyetracking systems rely on standard video cameras, which operate at frame rates of about 30 Hz. Such systems are limited to slow, full-frame operation. The video camera in the present system includes a charge-coupled-device (CCD) image detector plus electronic circuitry capable of implementing an advanced control scheme that effects readout from a small region of interest (ROI), or subwindow, of the full image. Inasmuch as the image features of interest (the cornea and pupil) typically occupy a small part of the camera frame, this ROI capability can be exploited to determine the direction of gaze at a high frame rate by reading out from the ROI that contains the cornea and pupil (but not from the rest of the image) repeatedly. One of the present algorithms exploits the ROI capability. The algorithm takes horizontal row slices and takes advantage of the symmetry of the pupil and cornea circles and of the gray-scale contrasts of the pupil and cornea with respect to other parts of the eye. The algorithm determines which horizontal image slices contain the pupil and cornea, and, on each valid slice, the end coordinates of the pupil and cornea. Information from multiple slices is then combined to robustly locate the centroids of the pupil and cornea images. The other of the two present algorithms is a modified version of an older algorithm for estimating the direction of gaze from the centroids of the pupil and cornea. The modification lies in the use of the coordinates of the centroids, rather than differences between the coordinates of the centroids, in a gaze-mapping equation. The equation locates a gaze point, defined as the intersection of the gaze axis with a surface of interest, which is typically a computer display screen (see figure). The expected advantage of the modification is to make the gaze computation less dependent on some simplifying assumptions that are sometimes not accurate

Slit-lamp exam

MedlinePlus

... eyes, especially the eyelids, cornea, conjunctiva, sclera, and iris. Often a yellow dye (fluorescein) is used to ... lens on the front of the eye) Eyelids Iris (colored part of the eye between the cornea ...

Broadly Applicable Nanowafer Drug Delivery System for Treating Eye Injuries

DTIC Science & Technology

2014-09-01

the drug molecular transport into the cornea. Intravital laser confocal imaging of the live mouse cornea demonstrating the presence of drug in the...vivo drug release in the mouse cornea by laser confocal fluorescence imaging study revealed that the nanowafers upon instillation on mouse eye were...C) 500nm; (D) 1µm; (E) 1.5µm; and (F) 3µm A B C D E F microscopy (SEM) for the feature integrity and uniformity. The SEM images revealed the presence

Excimer laser photorefractive surgery of the cornea

NASA Astrophysics Data System (ADS)

Gaster, Ronald N.

1998-09-01

The 193 nm argon fluoride (ArF) excimer laser can effectively be used to change the radius of curvature of the cornea and thus alter the refractive state of the eye. This change allows myopic (nearsighted) patients to see well with less dependence on glasses or contact lenses. The two major techniques of laser refractive surgery currently in effect in the United States are photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). This paper will discuss these refractive cornea surgical techniques.

C3 fixed in vivo to cornea from horses inoculated with Leptospira interrogans.

PubMed

Parma, A E; Cerone, S I; Sansinanea, S A; Ghezzi, M

1992-10-01

C3 was detected bound in vivo to the opaque cornea of horses inoculated with killed Leptospira interrogans. Employing epithelial corneal cells isolated from a monolayer in tissue culture, we proved that C3 is fixed in vitro to the intact cell surface after incubation with a fresh equine anti-Leptospira serum. These findings, in addition to the infiltration of cornea with neutrophils and lymphocytes, may explain the mechanisms of tissue damage in recurrent uveitis of horses with leptospirosis.

Blurry Worldview: Understanding Myopia

MedlinePlus

... the clearest possible vision. Eye surgery , such as LASIK and PRK, changes the shape of the cornea ... nearsighted or whose corneas are too thin for LASIK or PRK, are surgically implanted inside the eye. ...

Decreased Integrity, Content, and Increased Transcript Level of Mitochondrial DNA Are Associated with Keratoconus

PubMed Central

Hao, Xiao-Dan; Chen, Zhao-Li; Qu, Ming-Li; Zhao, Xiao-Wen; Li, Su-Xia; Chen, Peng

2016-01-01

Oxidative stress may play an important role in the pathogenesis of keratoconus (KC). Mitochondrial DNA (mtDNA) is involved in mitochondrial function, and the mtDNA content, integrity, and transcript level may affect the generation of reactive oxygen species (ROS) and be involved in the pathogenesis of KC. We designed a case-control study to research the relationship between KC and mtDNA integrity, content and transcription. One-hundred ninety-eight KC corneas and 106 normal corneas from Chinese patients were studied. Quantitative real-time PCR was used to measure the relative mtDNA content, transcript levels of mtDNA and related genes. Long-extension PCR was used to detect mtDNA damage. ROS, mitochondrial membrane potential and ATP were measured by respective assay kit, and Mito-Tracker Green was used to label the mitochondria. The relative mtDNA content of KC corneas was significantly lower than that of normal corneas (P = 9.19×10−24), possibly due to decreased expression of the mitochondrial transcription factor A (TFAM) gene (P = 3.26×10−3). In contrast, the transcript levels of mtDNA genes were significantly increased in KC corneas compared with normal corneas (NADH dehydrogenase subunit 1 [ND1]: P = 1.79×10−3; cytochrome c oxidase subunit 1 [COX1]: P = 1.54×10−3; NADH dehydrogenase subunit 1, [ND6]: P = 4.62×10−3). The latter may be the result of increased expression levels of mtDNA transcription-related genes mitochondrial RNA polymerase (POLRMT) (P = 2.55×10−4) and transcription factor B2 mitochondrial (TFB2M) (P = 7.88×10−5). KC corneas also had increased mtDNA damage (P = 3.63×10−10), higher ROS levels, and lower mitochondrial membrane potential and ATP levels compared with normal corneas. Decreased integrity, content and increased transcript level of mtDNA are associated with KC. These changes may affect the generation of ROS and play a role in the pathogenesis of KC. PMID:27783701

Contrasting cellular damage after Blue-IRIS and Femto-LASIK in cat cornea.

PubMed

Wozniak, Kaitlin T; Elkins, Noah; Brooks, Daniel R; Savage, Daniel E; MacRae, Scott; Ellis, Jonathan D; Knox, Wayne H; Huxlin, Krystel R

2017-12-01

Blue-intra-tissue refractive index shaping (Blue-IRIS) is a new approach to laser refractive correction of optical aberrations in the eye, which alters the refractive index of the cornea rather than changing its shape. Before it can be implemented in humans, it is critical to establish whether and to what extent, Blue-IRIS damages the cornea. Here, we contrasted the impact of -1.5 D cylinder refractive corrections inscribed using either Blue-IRIS or femtosecond laser in-situ keratomileusis (femto-LASIK) on corneal cell viability. Blue-IRIS was used to write a -1.5 D cylinder gradient index (GRIN) lens over a 2.5 mm by 2.5 mm area into the mid-stromal region of the cornea in six freshly-enucleated feline eyes. The same correction (-1.5 D cylinder) was inscribed into another four cat eyes using femto-LASIK. Six hours later, all corneas were processed for histology and stained for terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) and p-γ-H2AX to label damaged cells. In Blue-IRIS-treated corneas, no tissue was removed and TUNEL-stained cells were confined to the laser focal zone in the stroma. In femto-LASIK, photoablation removed 14 μm of anterior stroma, but in addition, TUNEL-positive cells clustered across the femto-flap, the epithelium at the flap edges and the stroma below the ablation zone. Keratocytes positive for p-γ-H2AX were seen adjacent to all Blue-IRIS focal zones, but were completely absent from femto-LASIK-treated corneas. Unlike femto-LASIK, Blue-IRIS attains refractive correction in the cornea without tissue removal and only causes minimal, localized keratocyte death within the laser focal zones. In addition, Blue-IRIS induced DNA modifications associated with phosphorylation of γ-H2AX in keratocytes adjacent to the laser focal zones. We posit that this p-γ-H2AX response is related to alterations in chromatin structure caused by localized changes in osmolarity, a possible mechanism for the induced refractive index changes. Copyright © 2017 Elsevier Ltd. All rights reserved.

Safety and Efficacy of OXB-202, a Genetically Engineered Tissue Therapy for the Prevention of Rejection in High-Risk Corneal Transplant Patients.

PubMed

Fouladi, Naghmeh; Parker, Maria; Kennedy, Vicky; Binley, Katie; McCloskey, Laura; Loader, Julie; Kelleher, Michelle; Mitrophanous, Kyriacos A; Stout, J Timothy; Ellis, Scott

2018-06-01

Due to both the avascularity of the cornea and the relatively immune-privileged status of the eye, corneal transplantation is one of the most successful clinical transplant procedures. However, in high-risk patients, which account for >20% of the 180,000 transplants carried out worldwide each year, the rejection rate is high due to vascularization of the recipient cornea. The main reason for graft failure is irreversible immunological rejection, and it is therefore unsurprising that neovascularization (NV; both pre and post grafting) is a significant risk factor for subsequent graft failure. NV is thus an attractive target to prevent corneal graft rejection. OXB-202 (previously known as EncorStat ® ) is a donor cornea modified prior to transplant by ex vivo genetic modification with genes encoding secretable forms of the angiostatic human proteins, endostatin and angiostatin. This is achieved using a lentiviral vector derived from the equine infectious anemia virus called pONYK1EiA, which subsequently prevents rejection by suppressing NV. Previously, it has been shown that rabbit donor corneas treated with pONYK1EiA substantially suppress corneal NV, opacity, and subsequent rejection in an aggressive rabbit model of cornea graft rejection. Here, efficacy data are presented in a second rabbit model, which more closely mirrors the clinical setting for high-risk corneal transplant patients, and safety data from a 3-month good laboratory practice toxicology and biodistribution study of pONYK1EiA-modified rabbit corneas in a rabbit corneal transplant model. It is shown that pONYK1EiA-modified rabbit corneas (OXB-202) significantly reduce corneal NV and the rate of corneal rejection in a dose-dependent fashion, and are tolerated with no adverse toxicological findings or significant biodistribution up to 13 weeks post surgery in these rabbit studies. In conclusion, angiogenesis is a valid target to prevent corneal graft rejection in a high-risk setting, and transplanted genetically modified corneas are safe and well-tolerated in an animal model. These data support the evaluation of OXB-202 in a first-in-human trial.

Cellular effects of mitomycin-C on human corneas after photorefractive keratectomy.

PubMed

Rajan, Madhavan S; O'Brart, David P S; Patmore, Anne; Marshall, John

2006-10-01

To investigate the effects of mitomycin-C (MMC) on epithelial and keratocyte cell kinetics after photorefractive keratectomy (PRK) using an in vitro human cornea model. Department of Academic Ophthalmology, Rayne Institute, St. Thomas' Hospital, London, United Kingdom. Twenty-four human eye-bank corneas were placed in a specially designed acrylic corneal holder and cultured using the air-interface organ culture technique for up to 4 weeks. The corneas were divided into 3 groups. Group 1 consisted of 8 human corneas that had -9.00 diopter (D) myopic PRK without MMC application. Group 2 consisted of 8 corneas that had -9.00 D PRK with MMC (0.2 microg/mL) application for 1 minute on the stromal surface after ablation. Group 3 consisted of 8 corneas that had -9.00 D PRK with 2-minute exposure to MMC (0.2 microg/mL). Temporal events in epithelial and keratocyte cell kinetics were evaluated using digital imaging, confocal microscopy, and light microscopy. Epithelial latency was significantly delayed with MMC application in Groups 2 and 3 (P<.001). Epithelial migration was delayed in Group 3 (2-minute exposure) compared to migration in Group 2 (P<.04), with a consequent delay in epithelial closure (P<.001). Group 3 corneas had poorly differentiated epithelium that was significantly thinner than in Groups 1 and 2 (P<.0001). A significant delay in keratocyte regeneration occurred after MMC application (P<.0005). At 4 weeks, the anterior stromal cell density was significantly lower in Group 3 than Group 2 (P<.001). There were no significant differences in the mid- and posterior stromal keratocyte density between the groups. Results suggest that epithelial healing after MMC is characterized by prolonged latency and decreased migration rate dependent on exposure time. Mitomycin C application did not result in increased loss of keratocytes, but it significantly delayed keratocyte repopulation in the anterior stroma. The use of MMC 0.2 microg/mL for 1 minute resulted in optimum modulation of healing characterized by reduced keratocyte activation with normal epithelial differentiation.

Type I Interferon and Lymphangiogenesis in the HSV-1 Infected Cornea – Are they Beneficial to the Host?

PubMed Central

Bryant-Hudson, Katie; Conrady, Christopher D.; Carr, Daniel J.J.

2013-01-01

Herpes simplex virus type 1 (HSV-1) is a highly successful pathogen that can result in significant human morbidity. Within the cornea, it was thought the initial recognition of the pathogen was through Toll-like receptors expressed on/in resident cells that then elicit pro-inflammatory cytokine production, activation of anti-viral pathways, and recruitment of leukocytes. However, our lab has uncovered a novel, TLR-independent innate sensor that supersedes TLR induction of anti-viral pathways following HSV-1 infection. In addition, we have also found HSV-1 induces the genesis of lymphatic vessels into the cornea proper by a mechanism independent of TLRs and unique in the field of neovascularization. This review will focus on these two innate immune events during acute HSV-1 infection of the cornea. PMID:23876483

Raman Microscopy: A Noninvasive Method to Visualize the Localizations of Biomolecules in the Cornea.

PubMed

Kaji, Yuichi; Akiyama, Toshihiro; Segawa, Hiroki; Oshika, Tetsuro; Kano, Hideaki

2017-11-01

In vivo and in situ visualization of biomolecules without pretreatment will be important for diagnosis and treatment of ocular disorders in the future. Recently, multiphoton microscopy, based on the nonlinear interactions between molecules and photons, has been applied to reveal the localizations of various molecules in tissues. We aimed to use multimodal multiphoton microscopy to visualize the localizations of specific biomolecules in rat corneas. Multiphoton images of the corneas were obtained from nonlinear signals of coherent anti-Stokes Raman scattering, third-order sum frequency generation, and second-harmonic generation. The localizations of the adhesion complex-containing basement membrane and Bowman layer were clearly visible in the third-order sum frequency generation images. The fine structure of type I collagen was observed in the corneal stroma in the second-harmonic generation images. The localizations of lipids, proteins, and nucleic acids (DNA/RNA) was obtained in the coherent anti-Stokes Raman scattering images. Imaging technologies have progressed significantly and been applied in medical fields. Optical coherence tomography and confocal microscopy are widely used but do not provide information on the molecular structure of the cornea. By contrast, multiphoton microscopy provides information on the molecular structure of living tissues. Using this technique, we successfully visualized the localizations of various biomolecules including lipids, proteins, and nucleic acids in the cornea. We speculate that multiphoton microscopy will provide essential information on the physiological and pathological conditions of the cornea, as well as molecular localizations in tissues without pretreatment.

Ultrasound-Enhanced Delivery of Antibiotics and Anti-Inflammatory Drugs into the Eye

PubMed Central

Nabili, Marjan; Patel, Hetal; Mahesh, Sankaranarayana P.; Liu, Ji; Geist, Craig; Zderic, Vesna

2013-01-01

Delivery of sufficient amounts of therapeutic drugs into the eye is often a challenging task. In this study, ultrasound application (frequencies of 400 KHz to 1 MHz, intensities of 0.3–1.0 W/cm2 and exposure duration of 5 min) was investigated to overcome the barrier properties of cornea, which is a typical route for topical administration of ophthalmic drugs. Permeability of ophthalmic drugs, tobramycin and dexamethasone and sodium fluorescein, a drug-mimicking compound, was studied in ultrasound- and sham-treated rabbit corneas in vitro using a standard diffusion cell setup. Light microscopy observations were used to determine ultrasound-induced structural changes in the cornea. For tobramycin, an increase in permeability for ultrasound- and sham-treated corneas was not statistically significant. Increase of 46%–126% and 32%–109% in corneal permeability was observed for sodium fluorescein and dexamethasone, respectively, with statistical significance (p < 0.05) achieved at all treatment parameter combinations (compared with sham treatments) except for 1-MHz ultrasound applications for dexamethasone experiments. This permeability increase was highest at 400 kHz and appeared to be higher at higher intensities applied. Histologic analysis showed structural changes that were limited to epithelial layers of cornea. In summary, ultrasound application provided enhancement of drug delivery, increasing the permeability of the cornea for the anti-inflammatory ocular drug dexamethasone. Future investigations are needed to determine the effectiveness and safety of this application in in vivo long-term survival studies. (E-mail: mnabili@gwu.edu) PMID:23415283

Corneal collagen cross-linking: a confocal, electron, and light microscopy study of eye bank corneas.

PubMed

Dhaliwal, Jasmeet S; Kaufman, Stephen C

2009-01-01

The purpose of this study was to evaluate morphological changes induced by corneal collagen cross-linking in a human ex vivo cornea, using confocal, electron, and light microscopy. The central epithelium was partially removed from ex vivo human corneal buttons. Riboflavin 0.1% solution was applied before ultraviolet A light treatment and then for every 2 minutes for 30 minutes while the corneas were exposed to ultraviolet A light at a wavelength of 370 nm and intensity of 3 mW/cm(2). Each cornea was evaluated using confocal, electron, and light microscopy. Confocal microscopy demonstrated normal-appearing corneas on their initial pretreatment examination, with reduced stromal detail. After treatment, a superficial layer of highly reflective spherical structures (4-10 microm) was observed. Many of these hyperreflective structures appeared up to a depth of 300 microm. The remainder of the corneal stroma and endothelium appeared normal. Electron microscopy showed keratocyte apoptotic changes to a depth of 300 microm. No observable pathologic changes were seen on histology. Based on clinical studies, corneal cross-linking is a promising treatment that appears to be safe and to halt ectatic corneal disease progression. Initial European studies used animal models to extrapolate human protocols. In conjunction with clinical studies, we believe that human ex vivo corneal studies provide a means to evaluate the structural and morphological changes associated with this procedure, within human corneas, in a manner that cannot be accomplished in vivo.

Corneal permeability for cement dust: prognosis for occupational safety

NASA Astrophysics Data System (ADS)

Kalmykov, R. V.; Popova, D. V.; Kamenskikh, T. G.; Genina, E. A.; Tuchin, V. V.; Bashkatov, A. N.

2018-02-01

The high dust content in air of a working zone causes prevalence of pathologies of the anterior segment of the eye of workers of cement production. Therefore, studying of features of cement dust impact on structure of a cornea and development of ways of eye protection from this influence is relevant. In this work experimental studies were carried out with twenty eyes of ten rabbits. OCTtomography was used to monitor the light attenuation coefficient of the cornea in vitro during the permeability of cement dust and/or keratoprotector (Systein Ultra). The permeability coefficients of the cornea for water, cement dust and keratoprotector were measured. A computer model allowing one to analyze the diffusion of these substances in the eye cornea was developed. It was shown that 1) the cement dust falling on the eye cornea caused pronounced dehydration of the tissue (thickness decreasing) and led to the increase of the attenuation coefficient, which could affect the deterioration of the eyesight of workers in the conditions of cement production; 2) the application of the keratoprotector to the eye cornea when exposed by cement dust, slowed significantly the dehydration process and did not cause the increase of the attenuation coefficient that characterized the stabilization of visual functions. At this, the keratoprotector itself did not cause dehydration and led to the decrease of the attenuation coefficient, which could allow it to be used for a long time in the order to protect the organ of vision from the negative effects of cement dust.

Ultraviolet photography of the in vivo human cornea unmasks the Hudson-Stähli line and physiologic vortex patterns.

PubMed

Every, Sean G; Leader, John P; Molteno, Anthony C B; Bevin, Tui H; Sanderson, Gordon

2005-10-01

To perform ultraviolet (UV) macrophotography of the normal in vivo human cornea, establishing biometric data of the major component of UV absorption for comparison with the Hudson-Stähli (HS) line, the distribution of iron demonstrated by Perl stain, and cases of typical amiodarone keratopathy. Nonrandomized comparative case series of UV photographs of 76 normal corneas (group 1) and 16 corneas with typical amiodarone keratopathy (group 2). Image-analysis software was used to grade the major component of UV absorption for slope and the coordinates of its points of intersection with the vertical corneal meridian and inflection. In group 1 the major component had a mean slope of 5.8 degrees, sloping down from nasal to temporal cornea. The mean coordinates of points of intersection with the vertical corneal meridian and inflection were (0, 0.30) and (0.02, 0.31), respectively. No significant differences between groups 1 and 2 were found for slope (P = 0.155), intersection with the vertical corneal meridian (P = 0.517), and point of inflection (P = 0.344). The major component of UV absorption was consistent with published characteristics of the HS line, and coincidence of UV absorption and Perl-stained iron was demonstrated in one corneal button. A vortex pattern of UV absorption was observed in all corneas. UV photography demonstrates subclinical corneal iron, confirming its deposition in an integrated HS line/vortex pattern. Coincident iron and amiodarone deposition occurs in amiodarone keratopathy.

Factors affecting DSAEK graft lenticle adhesion: an in vitro experimental study.

PubMed

Vaddavalli, Pravin K; Diakonis, Vasilios F; Canto, Ana P; Kankariya, Vardhaman P; Pappuru, Rajeev R; Ruggeri, Marco; Banitt, Michael R; Kymionis, George D; Yoo, Sonia H

2014-06-01

The aim of this study was to evaluate different factors that affect Descemet stripping automated endothelial keratoplasty (DSAEK) donor graft lenticle adhesion to the recipient cornea. This experimental study included 10 eye bank recipient corneas and 10 donor DSAEK lenticles. Recipient corneas were mounted on an artificial anterior chamber (AC), whereas donor lenticles were placed beneath the host cornea. Using optical coherence tomography and imaging software, the interface gap (IG) between the donor and recipient cornea was quantified to evaluate the effect of variations in AC air fill pressure, AC air fill duration, corneal massage, and corneal venting incisions on DSAEK donor graft lenticle adhesion. Different intraocular pressures (IOP) under air for the same time intervals, do not significantly correlate with the IG; nevertheless, it was noticed that the IG decreases as the IOP increases. With respect to the magnitude of AC IOP, there was no statistically significant difference when comparing 10 mm Hg with 30 mm Hg and assessing IG (P = 0.4). Complete air-fluid exchange resulted in significantly higher IG when compared with AC air bubble of 10 and 30 mm Hg that was sustained for 1 hour (P < 0.05). Furthermore, corneal surface massage did not facilitate DSAEK graft adhesion (P = 0.59). Finally, paracentral venting incisions followed by interface fluid aspiration seemed to significantly decrease the IG (P = 0.014). Corneal venting incisions and higher AC IOP values seem to facilitate DSAEK donor graft lenticle adhesion to the recipient cornea.

Sorption of sodium hydroxide by type I collagen and bovine corneas.

PubMed

Whikehart, D R; Edwards, W C; Pfister, R R

1991-01-01

There are no quantitative studies on the uptake of alkali into corneal tissues. To study this phenomenon, both type I collagen and bovine corneas were incubated in sodium hydroxide (NaOH) under varying conditions for periods up to 27.5 h. The sorption (absorption or adsorption) of the alkali to protein and tissue was measured as the quantity of NaOH no longer available for titration to neutrality with hydrochloric acid. Sorption was found to be dependent on the concentration of NaOH (0.01-1 N) but independent of the incubation temperature (4-35 degrees C). In whole cornea, sorption of 1 N NaOH began immediately and increased with time up to 6 h. After 6 h, sorption decreased, together with the observed degradation and solubilization of the tissue. Stripping of the corneal endothelium alone or of the endothelium and epithelium increased sorption in a similar manner when compared to whole corneas for periods up to 4 h. These observations are compatible with ionic and nonionic bonding of hydroxide ions to collagen (including that of the cornea) and the subsequent release of hydroxide ions during hydrolysis of the protein itself. Indirect evidence also suggests the inclusion of quantities of unbound hydroxide ions in hydrated gels of glycosaminoglycans. It is proposed that in a chemical burn of the cornea, alkali is both stored in the tissue (by sorption) and reacted with it (by hydrolysis), without any net consumption of alkali taking place.

The absorption characteristics of the human cornea in ultraviolet-a crosslinking.

PubMed

Koppen, Carina; Gobin, Laure; Tassignon, Marie-José

2010-03-01

With respect to the safety of ultraviolet-A (UVA) crosslinking for the corneal endothelium, an absorption coefficient is used that has been calculated in riboflavin soaked porcine corneas. We aim to validate this value for clinical use by measuring the absorption coefficient for UVA 365 nm in postmortem human corneas after instilling riboflavin on the corneal surface. Corneal thickness was measured in nine pairs of human donor eyes of which one eye was subjected to manual removal of the epithelium, whereas the epithelium of the fellow eye was left intact. Both eyes were instilled with riboflavin 0.1% in dextran 20% on the intact globe. After 20 min, the corneas were rinsed, and a corneoscleral button was trephined. The transmission of the cornea for UVA 365 nm was measured by transillumination, which allows calculation of the absorption coefficient. Measurement of average corneal thickness was 658.5 +/- 51.5 microm when the epithelium was removed, and 758.3 +/- 98.8 microm without epithelial removal. The average transmittance for UVA 365 nm was 12.89 +/- 4.10% with epithelial debridement and 28.52 +/- 4.39% without (P<0.05). The resultant average absorption coefficient is 32 +/- 5 cm when the epithelium is removed and 17 +/- 2 cm when it is left intact (P<0.05). Our results show an absorption coefficient for human corneas that is much lower than the values reported in the literature. This finding may be relevant when considering endothelial safety of the clinical crosslinking treatment.

[Evaluation of corneal tissue by slit lamp and specular microscopy during the preservation period].

PubMed

Farias, Roberta Jansen de Mello; Kubokawa, Karen Miyuki; Schirmer, Maurício; Sousa, Luciene Barbosa de

2007-01-01

To analyze changes evaluation of corneal grafts by slit lamp and electron microscopy examination in order to ensure a donor cornea of good quality level for the patient. 203 corneas donated to the Sorocaba Eye Bank between September 2002 and February 2003 were prospectively evaluated. The evaluation was graded from 0 to 3 according to the following factors: exposure and epithelial damage, stromal opacity, Descemet folds, pleomorphism, polymegatism and guttata, endothelial snail track, edema, reflectivity. The data were correlated with time between death and preservation, endothelial cell count and donor's age. 203 corneas were evaluated. The mean age of donors was 55 years (dp= 14.8 years). The mean time between death and preservation was 9.1 h (minimum of 2 h and maximum of 25 h). Eighty-six corneas suffered worsening of evaluation, whereby 66.3% had less than 2,500 endothelial cell count and 59.3% presented time between death and preservation over 6 hours. The mean day of the grading change was the 5th. Corneal grafts from younger donors had significantly better evaluation than those of older donors. The mean time of the evaluation change was on the 5th day in 50% of the corneas, however, an expressive number of corneas suffered changes on the 6th, 7th and 8th day. Corneal graft preserved after 6 hours of death had a greater tendency to lower cell count and to decrease in evaluation grades.

Intracellular Position of Centrioles and the Direction of Homeostatic Epithelial Cell Movements in the Mouse Cornea.

PubMed

Silverman, Erika; Zhao, Jin; Merriam, John C; Nagasaki, Takayuki

2017-02-01

Corneal epithelial cells exhibit continuous centripetal movements at a rate of about 30 µm per day, but neither the driving force nor the mechanism that determines the direction of movements is known. To facilitate the investigation of homeostatic cell movement, we examined if the intracellular position of a centriole can be used as a directional marker of epithelial cell movements in the mouse cornea. A direction of cell movements was estimated in fixed specimens from a pattern of underlying subepithelial nerve fibers. Intracellular position of centrioles was determined by gamma-tubulin immunohistology and plotted in a narrow strip along the entire diameter of a cornea from limbus to limbus. When we determined the position of centrioles in the peripheral cornea where cell movements proceed generally along a radial path, about 55% of basal epithelial cells contained a centriole in the front half of a cell. However, in the central cornea where cells exhibit a spiral pattern of movements, centrioles were distributed randomly. These results suggest that centrioles tend to be positioned toward the direction of movement in corneal basal epithelial cells when they are moving centripetally at a steady rate.

A Raman scattering and FT-IR spectroscopic study on the effect of the solar radiation in Antarctica on bovine cornea

NASA Astrophysics Data System (ADS)

Yamamoto, Tatsuyuki; Murakami, Naoki; Yoshikiyo, Keisuke; Takahashi, Tetsuya; Yamamoto, Naoyuki

2010-01-01

The Raman scattering and FT-IR spectra of the corneas, transported to the Syowa station in Antarctica and exposed to the solar radiation of the mid-summer for four weeks, were studied to reveal that type IV collagen involved in corneas were fragmented. The amide I and III Raman bands were observed at 1660 and 1245 cm -1, respectively, and the amide I and II infrared bands were observed at 1655 and 1545 cm -1, respectively, for original corneas before exposure. The background of Raman signals prominently increased and the ratio of amide II infrared band versus amide I decreased by the solar radiation in Antarctica. The control experiment using an artificial UV lamp was also performed in laboratory. The decline rate of the amide II/amide I was utilized for estimating the degree of fragmentation of collagen, to reveal that the addition of vitamin C suppressed the reaction while the addition of sugars promoted it. The effect of the solar radiation in Antarctica on the corneas was estimated as the same as the artificial UV lamp of four weeks (Raman) or one week (FT-IR) exposure.

Pretreatment methods to improve nerve immunostaining in corneas from long-term fixed embryonic quail eyes

NASA Technical Reports Server (NTRS)

Barrett, J. E.; Wells, D. C.; Conrad, G. W.

1999-01-01

Pretreatment methods were used to improve neurofilament immunostaining in corneas from embryonic day 16 Japanese quail corneas that had been stored in fixative solution for several months. A sequential combination of the following three pretreatments: brief microwave heating in saline, followed by extraction with sodium dodecyl sulfate (SDS) at 37 degrees C, followed by digestion with hyaluronidase at 37 degrees C, produced significantly increased antibody staining of corneal neurofilament proteins, compared with embryonic corneas subjected to no prior pretreatments or to single or two-step protocols. After applying the sequence of all three pretreatments, darkest nerve staining and increased numbers of fine branches were observed, together with lower background staining. Thus, the result of applying the three-step pretreatment sequence is better than that of applying any of its component single pretreatments or even combinations of any two of them. These findings therefore suggest that each of these three pretreatments causes a unique effect, beneficial to immunostaining of neurofilament proteins, and that their individual effects are independent and additive. In addition to embryonic corneas, the three-step procedure also may be useful for immunostaining of nerves in other very delicate, highly-hydrated tissues containing an abundance of extracellular matrix.

Morphological description of limbal epithelium: searching for stem cells crypts in the dog, cat, pig, cow, sheep and horse.

PubMed

Patruno, M; Perazzi, A; Martinello, T; Blaseotto, A; Di Iorio, E; Iacopetti, I

2017-06-01

The cornea provides protection and transparency to the eye, allowing an optimal sharpness view. In some pathological conditions the cornea is able to regenerate thanks to the presence of a stem cells reservoir present at the level of the transition area between cornea and sclera (limbus). Corneal cell therapies in Veterinary Medicine are really limited due to the lacking of knowledge about the anatomy of the limbal area, the putative presence of stem cells and their identification in domestic species. The aim of this study was to provide an overview of the main distinctive structural features of the sclero-corneal junction and conjunctival-corneal junction areas in some species of veterinary importance, using optic microscope observations of histological sections. The resulting data were compared with cornea from humans adapting protocols already used to identify stem cells by means of a specific cellular marker. We tested the expression of ΔNp63α isoform in the cornea basal cells, trying to correlate the distribution profile with areas of highly proliferative turnover. The results obtained from this study represent a first step towards the identification of a corneal stem cells reservoir in different animals.

Healing of corneal epithelial wounds in marine and freshwater fish.

PubMed

Ubels, J L; Edelhauser, H F

The corneal epithelium of a fish is in direct contact with the aquatic environment and is a barrier to movement of ions and water into and through the cornea. This tissue layer is thus important in maintenance of corneal transparency. When the epithelium is wounded, its protective function is lost and corneal transparency remains compromised until the epithelial barrier is re-established. This study was undertaken to investigate the healing response of the fish cornea to epithelial abrasion. Wounds were stained with fluorescein and photographed during healing. Wound areas were measured by planimetry. The cornea of the sculpin, a marine teleost, becomes edematous after wounding and heals at 2.54 to 3.42 mm2/hr. Nonswelling corneas of the elasmobranchs--dogfish shark and skate--heal at 1.29 mm2/hr, respectively. The wounded eye of the rainbow trout, a freshwater teleost, is stressed by the low osmolality of the environment. Severe corneal edema and cataracts develop following epithelial wounding, and the cornea heals at 0.64 mm2/hr. Although the healing rates in teleosts differ from those in mammals, histology shows that the corneal healing mechanism is essentially the same in fish and mammals.

Genetics Home Reference: congenital stromal corneal dystrophy

MedlinePlus

... decorin, which is involved in the organization of collagens. Collagens are proteins that strengthen and support connective tissues ... ligaments. In the cornea, well-organized bundles of collagen make the cornea transparent. Decorin ensures that collagen ...

Optical properties of an anterior lamellar human cornea model based on fibrin-agarose

NASA Astrophysics Data System (ADS)

Ionescu, Ana M.; Cardona, Juan de la Cruz; Ghinea, Razvan; Garzón, Ingrid; González-Andrades, Miguel; Alaminos, Miguel; Pérez, Maria del Mar

2017-08-01

The optical evaluation carried out using the Inverse Adding-Doubling (IAD) method to determine the scattering and the absorption coefficients of the bioengineered human corneal stromas showed that this type of artificial biomaterials shared many similarities with native control cornea after four weeks of development in culture. Their absorption and reduced scattering coefficients values were higher than the ones of the control cornea, but their spectral behaviors of both coefficients were similar. Time of development in culture was an influencing factor on the results.

Experience of Comamonas acidovorans keratitis with delayed onset and treatment response in immunocompromised cornea.

PubMed

Lee, Sang Mok; Kim, Mee Kum; Lee, Jae Lim; Wee, Won Ryang; Lee, Jin Hak

2008-03-01

To report 2 cases of Comamonas acidovorans keratitis in immunocompromised cornea. A complete review of the medical records of the two cases of Comamonas acidovorans keratitis. We found some similarities in clinical courses of two cases. Both of them showed development of keratitis during the management with corticosteroids, delayed onset, slow response to antibiotics, and relatively less affected corneal epithelium. Comamonas acidovorans is known as a less virulent organism. However it can cause an indolent infection that responds slowly even to adequate antibiotics therapy in immunocompromised corneas.

A fish eye out of water: epithelial surface projections on aerial and aquatic corneas of the 'four-eyed fish' Anableps anableps.

PubMed

Simmich, Joshua; Temple, Shelby E; Collin, Shaun P

2012-03-01

Vertebrate corneas feature a variety of microprojections, to which a tear film adheres. These microprojections are formed by folds in epithelial cell membranes, which increase surface area, stabilise the tear film and enhance movement of nutritional and waste products across cell membranes. Differences in corneal microprojections among vertebrates have been correlated with habitat and differ markedly between terrestrial and aquatic species. This study investigated epithelial microprojections of both the aerial (dorsal) and aquatic (ventral) corneal surfaces of the 'four-eyed fish' Anableps anableps using scanning electron microscopy. The central region of the dorsal cornea, which projects above the water, had a density of 16,387 ± 3,995 cells per mm(2) , while the central region of the ventral cornea (underwater) had a density of 22,428 ± 6,387 cells per mm(2), a difference that suggests an environmental adaptation along the two visual axes. Both corneal surfaces were found to possess microridges rather than microvilli or microplicae characteristic of terrestrial/aerial vertebrates. Microridges were 142 ± 9 nm wide and did not differ (p = 0.757) between dorsal and ventral corneas. Microridges were consistently separated by a distance of 369 ± 9 nm across both corneas. Dorsal-ventral differences in corneal epithelial cell density in Anableps anableps suggest a difference in osmotic pressure of the two corneas. The modest differences in the microprojections indicate that the need to secure the tear film underlying each optical axis is of prime importance, due to the likelihood that a persistent layer of water normally covers both dorsal and ventral corneal surfaces and that maintaining a transparent optical pathway for vision is critical for a species prone to predation from both above and below the water's surface. © 2012 The Authors. Clinical and Experimental Optometry © 2012 Optometrists Association Australia.

Lactoferrin Expression in Human and Murine Ocular Tissue.

PubMed

Rageh, Abrar A; Ferrington, Deborah A; Roehrich, Heidi; Yuan, Ching; Terluk, Marcia R; Nelson, Elizabeth F; Montezuma, Sandra R

2016-07-01

Lactoferrin (LF) is a multifunctional protein known to provide innate defense due to its antimicrobial and anti-inflammatory properties. In the eye, LF has been identified in the tears and vitreous humor. Its presence in other ocular tissues has not been determined. Our aim is to assess the presence of LF in the cornea, iris, retina and retinal pigment epithelium (RPE) of humans and mice. To test for the endogenous production of LF, reverse transcription polymerase chain reaction was performed in cultured human cells from the cornea and RPE and in murine tissues. To confirm LF localization in specific ocular tissue, immunohistochemistry was performed on flat mounts of cornea, retina and RPE in human donor eyes. The presence of LF was assessed by western blotting in human and mouse ocular tissue and human culture cells (cornea and RPE). To verify antibody specificity, purified human LF and transferrin (TF) were used on 1D and 2D western blots. LF gene expression was confirmed in the cornea and RPE cell cultures from humans, suggesting that LF is an endogenously produced protein. PCR results from mouse ocular tissue showed LF expression in cornea, iris, RPE, but not in retina. These results were also consistent with immunohistochemical localization of LF in human donor tissue. Antibody reaction for human LF was specific and western blotting showed its presence in the cornea, iris and RPE tissues. A faint reaction for the retina was observed but was likely due to contamination from other ocular tissues. Multiple commercially available antibodies for murine LF cross-reacted with TF, so no reliable results were obtained for murine western blot. LF is expressed in multiple eye tissues of humans and mice. This widespread expression and multifunctional activity of LF suggests that it may play an important role in protecting eye tissues from inflammation-associated diseases.

Effect of different culture media and deswelling agents on survival of human corneal endothelial and epithelial cells in vitro.

PubMed

Valtink, Monika; Donath, Patricia; Engelmann, Katrin; Knels, Lilla

2016-02-01

To examine the effects of media and deswelling agents on human corneal endothelial and epithelial cell viability using a previously developed screening system. The human corneal endothelial cell line HCEC-12 and the human corneal epithelial cell line HCE-T were cultured in four different corneal organ culture media (serum-supplemented: MEM +2 % FCS, CorneaMax®/CorneaJet®, serum-free: Human Endothelial-SFM, Stemalpha-2 and -3) with and without 6 % dextran T500 or 7 % HES 130/0.4. Standard growth media F99HCEC and DMEM/F12HCE-T served as controls. In additional controls, the stress inducers staurosporine or hydrogen peroxide were added. After 5 days in the test media, cell viability was assessed by flow cytometrically quantifying apoptotic and necrotic cells (sub-G1 DNA content, vital staining with YO-PRO-1® and propidium iodide) and intracellular reactive oxygen species (ROS). The MEM-based media were unable to support HCEC-12 and HCE-T survival under stress conditions, resulting in significantly increased numbers of apoptotic and necrotic cells. HCEC-12 survival was markedly improved in SFM-based media even under staurosporine or hydrogen peroxide. Likewise, HCE-T survival was improved in SFM with or without dextran. The media CorneaMax®, CorneaJet®, and CorneaMax® with HES supported HCEC-12 survival better than MEM-based media, but less well than SFM-based media. HCE-T viability was also supported by CorneaJet®, but not by CorneaMax® with or without HES. Stemalpha-based media were not suitable for maintaining viability of HCEC-12 or HCE-T in the applied cell culture system. The use of serum-supplemented MEM-based media for corneal organ culture should be discontinued in favour of serum-free media like SFM.

The management of cornea blindness from severe corneal scarring, with the Athens Protocol (transepithelial topography-guided PRK therapeutic remodeling, combined with same-day, collagen cross-linking).

PubMed

Kanellopoulos, Anastasios John

2012-01-01

To evaluate the safety and efficacy of combined transepithelial topography-guided photorefractive keratectomy (PRK) therapeutic remodeling, combined with same-day, collagen cross-linking (CXL). This protocol was used for the management of cornea blindness due to severe corneal scarring. A 57-year-old man had severe corneal blindness in both eyes. Both corneas had significant central scars attributed to a firework explosion 45 years ago, when the patient was 12 years old. Corrected distance visual acuity (CDVA) was 20/100 both eyes (OU) with refraction: +4.00, -4.50 at 135° in the right eye and +3.50, -1.00 at 55° in the left. Respective keratometries were: 42.3, 60.4 at 17° and 35.8, 39.1 at 151.3°. Cornea transplantation was the recommendation by multiple cornea specialists as the treatment of choice. We decided prior to considering a transplant to employ the Athens Protocol (combined topography-guided partial PRK and CXL) in the right eye in February 2010 and in the left eye in September 2010. The treatment plan for both eyes was designed on the topography-guided wavelight excimer laser platform. Fifteen months after the right eye treatment, the right cornea had improved translucency and was topographically stable with uncorrected distance visual acuity (UDVA) 20/50 and CDVA 20/40 with refraction +0.50, -2.00 at 5°. We noted a similar outcome after similar treatment applied in the left eye with UDVA 20/50 and CDVA 20/40 with -0.50, -2.00 at 170° at the 8-month follow-up. In this case, the introduction of successful management of severe cornea abnormalities and scarring with the Athens Protocol may provide an effective alternative to other existing surgical or medical options.

The management of cornea blindness from severe corneal scarring, with the Athens Protocol (transepithelial topography-guided PRK therapeutic remodeling, combined with same-day, collagen cross-linking)

PubMed Central

Kanellopoulos, Anastasios John

2012-01-01

Purpose To evaluate the safety and efficacy of combined transepithelial topography-guided photorefractive keratectomy (PRK) therapeutic remodeling, combined with same-day, collagen cross-linking (CXL). This protocol was used for the management of cornea blindness due to severe corneal scarring. Methods A 57-year-old man had severe corneal blindness in both eyes. Both corneas had significant central scars attributed to a firework explosion 45 years ago, when the patient was 12 years old. Corrected distance visual acuity (CDVA) was 20/100 both eyes (OU) with refraction: +4.00, −4.50 at 135° in the right eye and +3.50, −1.00 at 55° in the left. Respective keratometries were: 42.3, 60.4 at 17° and 35.8, 39.1 at 151.3°. Cornea transplantation was the recommendation by multiple cornea specialists as the treatment of choice. We decided prior to considering a transplant to employ the Athens Protocol (combined topography-guided partial PRK and CXL) in the right eye in February 2010 and in the left eye in September 2010. The treatment plan for both eyes was designed on the topography-guided wavelight excimer laser platform. Results Fifteen months after the right eye treatment, the right cornea had improved translucency and was topographically stable with uncorrected distance visual acuity (UDVA) 20/50 and CDVA 20/40 with refraction +0.50, −2.00 at 5°. We noted a similar outcome after similar treatment applied in the left eye with UDVA 20/50 and CDVA 20/40 with −0.50, −2.00 at 170° at the 8-month follow-up. Conclusion In this case, the introduction of successful management of severe cornea abnormalities and scarring with the Athens Protocol may provide an effective alternative to other existing surgical or medical options. PMID:22347790

Bee sting of the cornea: a case study and review of the literature.

PubMed

Caça, Ihsan; Ari, Seyhmus; Ulü, Kaan; Ayata, Ali

2006-01-01

Bee stings of the cornea are rarely reported, but have the potential for causing serious ophthalmological injuries. We present a case of corneal bee sting with retained stinger apparatus. A 35-year-old patient presented with an acute, corneal bee sting of the right eye 12 hours after he was stung. The patient suffered from pain, blurred vision, and epiphora. The right eye showed edema of the upper and lower eyelid, conjunctival hyperemia, chemosis, and striate keratitis of the paracentral cornea by biomicroscopic examination. The stinger was identified in the depth of the corneal infiltration. Visual acuity was 5/10. It was removed surgically. After 2 months, the eye only showed a minimal residual corneal opacification. Visual acuity was 10/10. We present a case of bee sting to the cornea with retained stinger apparatus and treatment of this unusual presentation.

Intra-Ocular Pressure Measurement in a Patient with a Thin, Thick or Abnormal Cornea.

PubMed

Clement, Colin I; Parker, Douglas G A; Goldberg, Ivan

2016-01-01

Accurate measurement of intra-ocular pressure is a fundamental component of the ocular examination. The most common method of measuring IOP is by Goldmann applanation tonometry, the accuracy of which is influenced by the thickness and biomechanical properties of the cornea. Algorithms devised to correct for corneal thickness to estimate IOP oversimplify the effects of corneal biomechanics. The viscous and elastic properties of the cornea influence IOP measurements in unpredictable ways, a finding borne out in studies of patients with inherently abnormal and surgically altered corneal biomechanics. Dynamic contour tonometry, rebound tonometry and the ocular response analyzer provide useful alternatives to GAT in patients with abnormal corneas, such as those who have undergone laser vision correction or keratoplasty. This article reviews the various methods of intra-ocular pressure measurement available to the clinician and the ways in which their utility is influenced by variations in corneal thickness and biomechanics.

Mass Spectrometric Analyses of Phosphatidylcholines in Alkali-Exposed Corneal Tissue

PubMed Central

Crane, Ashley M.; Hua, Hong-Uyen; Coggin, Andrew D.; Gugiu, Bogdan G.; Lam, Byron L.; Bhattacharya, Sanjoy K.

2012-01-01

Purpose. The aims were to determine whether exposure to sodium hydroxide results in predictable changes in phosphatidylcholine (PC) in corneal tissue and if PC profile changes correlate to exposure duration. PCs are major components of the cell membrane lipid bilayer and are often involved in biological processes such as signaling. Methods. Enucleated porcine (n = 140) and cadaver human eyes (n = 20) were exposed to water (control) and 11 M NaOH. The corneas were excised and lipids were extracted using the Bligh and Dyer method with suitable modifications. Class-specific lipid identification was carried out using a ratiometric lipid standard on a TSQ Quantum Access Max mass spectrometer. Protein amounts were determined using Bradford assays. Results. Control and alkali-treated corneas showed reproducible PC spectra for both porcine and human corneas. Over 200 PCs were identified for human and porcine control and each experimental time point. Several PC species (m/z values) consequent upon alkali exposure could not be ascribed to a recorded PC species. Control and treated groups showed 41 and 29 common species among them for porcine and human corneas, respectively. The unique PC species peaked at 12 minutes and at 30 minutes for human and porcine corneas followed by a decline consistent with an interplay of alkali penetration and hydrolyses at various time points. Conclusions. Alkali exposure dramatically changes the PC profile of cornea. Our data are consistent with penetration and hydrolysis as stochastic contributors to changes in PCs due to exposure to alkali for a finite duration and amount. PMID:22956606

Dynamic OCT measurement of corneal deformation by an air puff in normal and cross-linked corneas

PubMed Central

Dorronsoro, Carlos; Pascual, Daniel; Pérez-Merino, Pablo; Kling, Sabine; Marcos, Susana

2012-01-01

A new technique is presented for the non-invasive imaging of the dynamic response of the cornea to an air puff inducing a deformation. 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. The entire deformation process can be dynamically visualized. A quantitative analysis allows direct extraction of several deformation parameters, such as amplitude, diameter and volume of the maximum deformation, as well as duration and speed of the increasing deformation period and the recovery period. The potential of the technique is demonstrated on porcine corneas in vitro under constant IOP for several conditions (untreated, after riboflavin instillation and under cross-linking with ultraviolet light), as well as on human corneas in vivo. The new technique has proved very sensitive to detect differences in the deformation parameters across conditions. We have confirmed non-invasively that Riboflavin and UV-cross-linking induce changes in the corneal biomechanical properties. Those differences appear to be the result of changes in constituent properties of the cornea, and not a consequence of changes in corneal thickness, geometry or IOP. These measurements are a first step for the estimation of the biomechanical properties of corneal tissue, at an individual level and in vivo, to improve diagnosis and prognosis of diseases and treatments involving changes in the biomechanical properties of the cornea. PMID:22435096

The assessment of the stability of the corneal structure after LASIK correction of myopia by different optical zone diameters.

PubMed

Milivojević, Milorad; Petrović, Vladimir; Vukosavljević, Miroslav; Marjanović, Ivan; Resan, Mirko

2016-06-01

Enlargement of optical zone (OZ) diameter during laser in situ keratomileusis (LASIK) correction of myopia postoperatively improves the optical outcome, however, it also leads to the increased stroma tissue consumption--progressive corneal thinning. The aim of this investigation was to present the possibility of safe OZ enlargement without impairing the structural stability of the cornea, while obtaining an improved optical outcome with LASIK treatment of short-sightedness. Preoperative assessment of the cornea structure and prediction of the ablated stroma tissue consumption was conducted in 37 patients (74 eyes) treated for short-sightedness by means of the LASIK method. With the eyes that, according to their cornea structure, had the capacity for OZ diameter enlargement of 0.5 mm, LASIK treatment was performed within the wider OZ diameter of 7.0 mm compared to the standard 6.5 mm. The following two groups were formed, depending on the diameter of the utilized OZ: the group I (the eyes treated with the OZ 6.5 mm, n = 37) and the group II (the eyes treated with the OZ 7.0 mm, n = 37). No significant difference in the observed structural parameters of the cornea was detected between the groups of patients treated with different OZ diameters. The values of all the parameters were significantly bellow the threshold values for the development of postoperative ectasia. Diameter enlargement of the treated OZ, if there is a preoperative cornea capacity for such enlargement, will not impair the postoperative stability of the cornea structure, and will significantly improve the optical outcome.

In vivo three-dimensional confocal laser scanning microscopy of the epithelial nerve structure in the human cornea.

PubMed

Stachs, Oliver; Zhivov, Andrey; Kraak, Robert; Stave, Joachim; Guthoff, Rudolf

2007-04-01

Evaluation of a new method for in vivo visualization of the distribution and morphology of human anterior corneal nerves. The anterior cornea was examined to a depth of 100 microm in four human volunteers with a confocal laser scanning microscope (CLSM) using a Rostock Cornea Module (developed in house) attached to a Heidelberg Retina Tomograph II (Heidelberg Engineering, Germany). Optical sections were digitally reconstructed in 3D using AMIRA (TGS Inc., USA). The scanned volumes had a greatest size of 300 x 300 x 40 microm and voxel size of 0.78 x 0.78 x 0.95 microm. The spatial arrangement of the epithelium, nerves and keratocytes was visualized by in vivo 3D-CLSM. The 3D-reconstruction of the volunteers' corneas in combination with the oblique sections gave a picture of the nerves in the central human cornea. Thin nerves run in the subepithelial plexus aligned parallel to Bowman's layer and are partially interconnected. The diameter of these fibres varied between 1.0 and 5 microm. Thick fibres rose out of the deeper stroma. The diameter of the main nerve trunks was 12+/-2 microm. Branches penetrating the anterior epithelial cell layer could not be visualized. 3D-CLSM allows analysis of the spatial arrangement of the anterior corneal nerves and visualization of the epithelium and keratocytes in the living human cornea. The developed method provides a basis for further studies of alterations of the cellular arrangement and epithelial innervation in corneal disease. This may help to clarify alterations of nerve fibre patterns under various clinical and experimental conditions.

Quantified histopathology of the keratoconic cornea.

PubMed

Mathew, Jessica H; Goosey, John D; Bergmanson, Jan P G

2011-08-01

This study systematically investigated and quantified histopathological changes in a series of keratoconic (Kc) corneas using a physiologically formulated fixative to not further distort the already distorted diseased corneas. Twelve surgically removed Kc corneal buttons were immediately preserved and processed for light and transmission electron microscopy using an established corneal protocol. Measurements were taken from the central cone and peripheral regions of the host button. The sample size examined ranged in length from 390 to 2608 μm centrally and 439 to 2242 μm peripherally. The average corneal thickness was 437 μm centrally and 559 μm peripherally. Epithelial thickness varied centrally from 14 to 92 μm and peripherally from 30 to 91 μm. A marked thickening of the epithelial basement membrane was noted in 58% of corneas. Centrally, anterior limiting lamina (ALL) was thinned or lost over 60% of the area examined, whereas peripheral cornea was also affected but to a lesser extent. Histopathologically, posterior cornea remained undisturbed by the disease. Anteriorly in the stroma, an increased number of cells and tissue debris were encountered, and some of these cells were clearly not keratocytes. It is concluded that Kc pathology, at least initially, has a distinct anterior focus involving the epithelium, ALL, and anterior stroma. The epithelium had lost its cellular uniformity and was compromised by the loss or damage to the ALL. The activity of the hitherto unreported recruited stromal cells may be to break down and remove ALL and anterior stromal lamellae, leading to the overall thinning that accompanies this disease.

Factors influencing the contamination rate of human organ-cultured corneas.

PubMed

Röck, Daniel; Wude, Johanna; Bartz-Schmidt, Karl U; Yoeruek, Efdal; Thaler, Sebastian; Röck, Tobias

2017-12-01

To assess the influence of donor, environment and storage factors on the contamination rate of organ-cultured corneas, to consider the microbiological species causing corneal contamination and to investigate the corresponding sensitivities. Data from 1340 consecutive donor corneas were analysed retrospectively. Logistic regression analysis was used to assess the influence of different factors on the contamination rate of organ-cultured corneas for transplantation. The mean annual contamination rate was 1.8 ± 0.4% (range: 1.3-2.1%); 50% contaminations were of fungal origin with exclusively Candida species, and 50% contaminations were of bacterial origin with Staphylococcus species being predominant. The cause of donor death including infection and multiple organ dysfunction syndrome increased the risk of bacterial or fungal contamination during organ culture (p = 0.007 and p = 0.014, respectively). Differentiating between septic and aseptic donors showed an increased risk of contamination for septic donors (p = 0.0020). Mean monthly temperature including warmer months increased the risk of contamination significantly (p = 0.0031). Sex, donor age, death to enucleation, death to corneoscleral disc excision and storage time did not increase the risk of contamination significantly. The genesis of microbial contamination in organ-cultured donor corneas seems to be multifactorial. The main source of fungal or bacterial contamination could be resident species from the skin flora. The rate of microbial contamination in organ-cultured donor corneas seems to be dependent on the cause of donor death and mean monthly temperature. © 2017 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Effects of vitamin D receptor knockout on cornea epithelium gap junctions.

PubMed

Lu, Xiaowen; Watsky, Mitchell A

2014-05-06

Gap junctions are present in all corneal cell types and have been shown to have a critical role in cell phenotype determination. Vitamin D has been shown to influence cell differentiation, and recent work demonstrates the presence of vitamin D in the ocular anterior segment. This study measured and compared gap junction diffusion coefficients among different cornea epithelium phenotypes and in keratocytes using a noninvasive technique, fluorescence recovery after photobleaching (FRAP), and examined the influence of vitamin D receptor (VDR) knockout on epithelial gap junction communication in intact corneas. Previous gap junction studies in cornea epithelium and keratocytes were performed using cultured cells or ex vivo invasive techniques. These invasive techniques were unable to measure diffusion coefficients and likely were disruptive to normal cell physiology. Corneas from VDR knockout and control mice were stained with 5(6)-carboxyfluorescein diacetate (CFDA). Gap junction diffusion coefficients of the corneal epithelium phenotypes and of keratocytes, residing in intact corneas, were detected using FRAP. Diffusion coefficients equaled 18.7, 9.8, 5.6, and 4.2 μm(2)/s for superficial squamous cells, middle wing cells, basal cells, and keratocytes, respectively. Corneal thickness, superficial cell size, and the superficial squamous cell diffusion coefficient of 10-week-old VDR knockout mice were significantly lower than those of control mice (P < 0.01). The superficial cell diffusion coefficient of heterozygous mice was significantly lower than control mice (P < 0.05). Our results demonstrate differences in gap junction dye spread among the epithelial cell phenotypes, mirroring the epithelial developmental axis. The VDR knockout influences previously unreported cell-to-cell communication in superficial epithelium.

Ultrasound-enhanced delivery of antibiotics and anti-inflammatory drugs into the eye.

PubMed

Nabili, Marjan; Patel, Hetal; Mahesh, Sankaranarayana P; Liu, Ji; Geist, Craig; Zderic, Vesna

2013-04-01

Delivery of sufficient amounts of therapeutic drugs into the eye is often a challenging task. In this study, ultrasound application (frequencies of 400 KHz to 1 MHz, intensities of 0.3-1.0 W/cm(2) and exposure duration of 5 min) was investigated to overcome the barrier properties of cornea, which is a typical route for topical administration of ophthalmic drugs. Permeability of ophthalmic drugs, tobramycin and dexamethasone and sodium fluorescein, a drug-mimicking compound, was studied in ultrasound- and sham-treated rabbit corneas in vitro using a standard diffusion cell setup. Light microscopy observations were used to determine ultrasound-induced structural changes in the cornea. For tobramycin, an increase in permeability for ultrasound- and sham-treated corneas was not statistically significant. Increase of 46%-126% and 32%-109% in corneal permeability was observed for sodium fluorescein and dexamethasone, respectively, with statistical significance (p < 0.05) achieved at all treatment parameter combinations (compared with sham treatments) except for 1-MHz ultrasound applications for dexamethasone experiments. This permeability increase was highest at 400 kHz and appeared to be higher at higher intensities applied. Histologic analysis showed structural changes that were limited to epithelial layers of cornea. In summary, ultrasound application provided enhancement of drug delivery, increasing the permeability of the cornea for the anti-inflammatory ocular drug dexamethasone. Future investigations are needed to determine the effectiveness and safety of this application in in vivo long-term survival studies. Copyright © 2013 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

Subclinical HSV-1 infections provide site-specific resistance to an unrelated pathogen

PubMed Central

Rowe, Alexander M.; Yun, Hongming; Treat, Benjamin R.; Kinchington, Paul R.; Hendricks, Robert L.

2016-01-01

Herpes Simplex Virus-1 (HSV-1) infections of the cornea range in severity from minor transient discomfort to the blinding disease Herpes Stromal Keratitis (HSK), yet most patients experience a single episode of epithelial keratitis followed by reestablishment of a clear cornea. We asked if a single transient episode of HSV-1 epithelial keratitis causes long-term changes in the corneal microenvironment that influence immune responses to subsequent corneal infection or trauma. We showed that C57BL/6 mouse corneas infected with HSV-1 KOS, which induces transient herpes epithelial keratitis without HSK sequelae, possessed a significant leukocytic infiltrate comprised primarily of CD4+ T cells and macrophages along with elevated chemokines and cytokines that persisted without loss of corneal clarity (subclinical inflammation). Chemokine and cytokine expression was CD4+T cell-dependent, in that their production was significantly reduced by systemic CD4+T cell depletion starting before infection, although short-term (3 day) local CD4+ T cell depletion after infection did not influence chemokine levels in cornea. Corneas with subclinical inflammation developed significantly greater trauma-induced inflammation when they were recipients of syngeneic corneal transplants, but also exhibited significantly increased resistance to infections by unrelated pathogens such as pseudorabies virus (PRV). The resistance to PRV was CD4+ T cell-dependent, since it was eliminated by local CD4+T cell-depletion from the cornea. We conclude that transient HSV-1 corneal infections cause long-term alterations of the corneal microenvironment that provide CD4-dependent innate resistance to subsequent infections by antigenically unrelated pathogens. PMID:28062697

The influence of Australian eye banking practices on corneal graft survival.

PubMed

Keane, Miriam C; Lowe, Marie T; Coster, Douglas J; Pollock, Graeme A; Williams, Keryn A

2013-08-19

To identify eye banking practices that influence corneal graft survival. Prospective cohort study of records of 19,254 followed corneal grafts in 15160 patients, submitted to the Australian Corneal Graft Registry between May 1985 and July 2012. Influence of corneal preservation method (organ culture, moist pot, Optisol, other); death-to-enucleation, death-to-preservation and enucleation-to-graft times; transportation by air; graft era; and indication for graft on probability of graft survival at most recent follow-up. In multivariate analysis, 919 penetrating grafts performed using corneas transported interstate by air exhibited worse survival than 14,684 grafts performed using corneas retrieved and used locally (hazard ratio [HR], 1.44; 95% CI, 1.21-1.73; P = 0.001). This was also the case for traditional lamellar grafts (64 corneas transported by air and 813 used locally; HR, 1.69; 95% CI, 1.03-2.78; P = 0.038). Indication for graft influenced survival of penetrating grafts (4611 keratoconus, 727 emergency or high-risk, 10,265 other indication; global P < 0.001) and traditional lamellar grafts (65 keratoconus, 212 emergency or high-risk, 600 other indication; global P < 0.001). The preservation medium in which corneas used for traditional lamellar grafts were stored exerted a marginal influence on graft survival (global P = 0.047). Donor corneas transported interstate exhibited poorer survival after transplantation than those retrieved and grafted locally. Higher proportions of emergency procedures involving transported corneas did not account for this difference. Where possible, efforts to avoid transportation of corneal tissue by air freight within Australia may be warranted.

[The cornea: stasis and dynamics].

PubMed

Nishida, Teruo

2008-03-01

The physiological roles of the cornea are to conduct external light into the eye, focus it, together with the lens, onto the retina, and to provide rigidity to the entire eyeball. Good vision thus requires maintenance of the transparency and proper refractive shape of the cornea. Although the cornea appears to be a relatively static structure, dynamic processes operate within and around the cornea at the tissue, cell, and molecular level. In this article, I review the mechanisms responsible for maintenance of corneal homeostasis as well as the development of new modes of treatment for various corneal diseases. I. The static cornea: structure and physiological functions. The cornea is derived from ectoderm, so that it can be considered as transparent skin. It is devoid of blood vessels and manifests the highest sensitivity in the entire body. The surface of the cornea is covered by tear fluid, which serves both as a lubricant and as a conduit for regulatory molecules. The cornea is also supplied with oxygen and various nutrients by the aqueous humor and a loop vascular system in addition to tear fluid. The cornea interacts with its surrounding tissues directly as well as indirectly through tear fluid or aqueous humor, with such interactions playing an important role in the regulation of corneal structure and functions. The resident cells of the cornea-epithelial cells, fibroblasts (keratocytes), and endothelial cells--also engage in mutual interactions through network systems. These interactions as well as those with infiltrated cells and regulation by nerves contribute to the maintenance of the normal structure and functions of the cornea as well as to the repair of corneal injuries. II. The dynamic cornea: maintenance of structure and functions by network systems. Developments in laser and computer technology have allowed observation of the cells and collagen fibers within the cornea. Furthermore, progress in cell and molecular biology has allowed characterization of dynamic network systems-including cell-cell and cell-extracellular matrix interactions as well as cytokines and neural factors-that contribute to the maintenance of corneal transparency and shape. III. Disruption of network systems: persistent corneal epithelial defects and corneal ulcer. Selection of the appropriate treatment for pathologic lesions of the cornea and the accompanying decrease in visual acuity requires localization of the lesion with regard to the epithelium, stroma, or endothelium of the cornea. In certain instances, however, it is not possible to determine the cause of the problem within the cornea. In such cases, the cause of the pathologic lesion and the target for treatment may lie in the surrounding tissues or environment. For example, corneal epithelial wound healing may be delayed, leading to the development of persistent epithelial defects, as a result of disruption of intercellular junctions between epithelial cells, an abnormality of the corneal basement membrane, altered concentrations of various cytokines in tear fluid, a lowered corneal sensation, or allergic reactions in the lid conjunctiva. Loss of corneal epithelial barrier function can further allow inflammatory cytokines present in tear fluid, together with infiltrated cells, to activate keratocytes and elicit excessive degradation of collagen in the stroma, thereby giving rise to corneal ulcer. IV. Development of new drugs for corneal diseases. We have attempted to apply the results of basic scientific research to the development of new drugs for corneal diseases that remain difficult to treat. The process of authorization for new drugs from the Ministry of Health, Labor, and Welfare takes more than two decades, however. The path from the bench to clinical practice is thus a long one. 1. Development of eyedrops for treatment of persistent corneal epithelial defects. We demonstrated the clinical efficacy of fibronectin eyedrops for the treatment of persistent epithelial defects of the cornea. However, the possibility of blood-borne infections has interfered with the development of serum-derived fibronectin as a drug. An automated machine for the preparation of autologous fibronectin eyedrops has therefore recently been developed. Furthermore, in seeking an alternative to fibronectin eyedrops, we are investigating the effects of a peptide corresponding to the second cell-binding domain of fibronectin on corneal epithelial wound healing. Considering that urokinase-type plasminogen activator may be expressed at the site of corneal epithelial defects and facilitates epithelial migration, the potential clinical application of annexin V, which stimulates the secretion of urokinase-type plasminogen activator for the treatment of persistent corneal epithelial defects is also now under investigation in Japan. 2. Development of eyedrops for treatment of neurotrophic keratopathy. Substance P, a neurotransmitter, stimulates corneal epithelial migration in a synergistic manner with insulin-like growth factor (IGF)--1. We have shown that eyedrops containing both the substance P-derived peptide FGLM-amide and the IGF-1--derived peptide SSSR are effective for the treatment of persistent corneal epithelial defects in individuals with diabetic keratopathy or neurotrophic keratopathy, both of which are associated with a reduction in corneal sensation. 3. Development of drugs for corneal ulcer. Treatment of corneal infection with antibiotics does not necessarily halt the process of corneal ulceration, which is characterized by excessive degradation of stromal collagen, or resolve persistent corneal epithelial defects. In addition to eyedrops for the treatment of persistent corneal epithelial defects, we have therefore also been working on the development of new drugs for the treatment of corneal ulcer. To this end, we have established an experimental system in which corneal fibroblasts are cultured in a three-dimensional collagen gel. With this system, we have shown that triptolide and steroids inhibit collagen degradation by corneal fibroblasts. Triptolide or its derivatives are thus potential drugs for the treatment of corneal ulcer and would work by acting directly on corneal fibroblasts rather than by inhibiting the secreted enzymes(matrix metalloproteinases) responsible for collagen degradation.

Sclera

MedlinePlus

... clear front section of the eye) to the optic nerve at the back of the eye. The sclera gives the eyeball its white color. The cornea and sclera are made of the same type of collagen fibers. In the cornea, the fibers are arranged in ...

Specific demographic factors could predict deceased potential cornea donors: A retrospective study from Beijing Tongren Hospital Eye Bank.

PubMed

Liu, Yang; Zhang, Yingnan; Zhang, Jing; Yu, Jin; Liang, Qingfeng; Pan, Zhiqiang

2017-12-01

Compared with evident cornea donors (ECDs), deceased potential cornea donors (DPCDs) have no obvious donor identifications to reference, which causes many eligible cornea tissues to be wasted. The demographic characteristics of DPCDs might be different from those of ECDs owing to the following different features: donation consent provided by relatives and willingness to donate before death. Thus, the aim of this study is to reveal the demographic characteristics of DPCDs by comparing DPCDs and ECDs.The demographic factors of 138 donors (both DPCDs and ECDs) were collected from the Beijing Tongren Hospital Eye Bank database and analyzed. To differentiate DPCDs from ECDs using the above-mentioned features, we interviewed the relatives of the donors by telephone. The relatives' attitudes toward cornea donation and their suggestions for our donation service were also acquired during the interview. Two logistic regressions were performed to reveal the demographic factors influencing the 2 features and indicate DPCDs.The donors had certain demographic characteristics (elderly, secondary, or tertiary education level, central district resident), and the most frequent cause of death for the donors was a malignant tumor (n = 56, 43.1%). All the relatives had positive attitudes toward cornea donations, and they hoped to increase publicity efforts to encourage more people to donate and establish more convenient and efficient access for cornea donation. In univariate regressions, age (P = .004, >50 years: odds ratio [OR] = 6.89, 95% confidence interval [CI]: 1.82-26.05), marital status (P = .043, divorced: OR = 9.00,95% CI: 1.33-60.80) significantly influenced relative consent, whereas age (P = .001, >50 years: OR = 15.00, 95% CI: 3.00-74.98), and family address (P = .001, central district: OR = 1) were significant factors influencing the willingness to donate before death. In multivariate regression, age (P = .021, >50 years: OR = 8.14, 95% CI: 1.37-48.41) was the only significant factor influencing relative consent. Similarly, age (P = .02, >50 years: OR = 7.55, 95% CI: 1.21-47.25) was the only factor influencing willingness to donate before death.In conclusion, specific demographic factors could indicate DPCDs and might reveal directions and methods for cornea donation coordination in the future.

Descemet membrane adhesion strength is greater in diabetics with advanced disease compared to healthy donor corneas.

PubMed

Schwarz, Chaid; Aldrich, Benjamin T; Burckart, Kimberlee A; Schmidt, Gregory A; Zimmerman, M Bridget; Reed, Cynthia R; Greiner, Mark A; Sander, Edward A

2016-12-01

Descemet membrane endothelial keratoplasty (DMEK) is an increasingly popular surgical procedure for treating ocular diseases that require a corneal transplant. Previous studies have found that tissue tearing during surgical preparation is more likely elevated in eyes from donors with a history of diabetes mellitus. To quantify these potential differences, we established an experimental technique for quantifying the force required to separate the endothelium-Descemet membrane complex (EDM) from stroma in human donor corneal tissue, and we assessed differences in adhesion strength between diabetic and non-diabetic donor corneas. Transplant suitable corneas were obtained from 23 donors 50-75 years old with an average preservation to assay time of 11.5 days. Corneas were classified from a medical records review as non-diabetic (ND, n = 9), diabetic without evidence of advanced disease (NAD, n = 8), or diabetic with evidence of advanced disease (AD, n = 10). Corneas were sectioned into 3 mm wide strips and the EDM peeled from the stroma. Using the force-extension data obtained from mechanical peel testing, EDM elastic peel tension (T E ), elastic stiffness (S E ), average delamination tension (T D ), and maximum tension (T MAX ) were calculated. Mean T E , S E , T D , and T MAX values for ND corneas were 0.78 ± 0.07 mN/mm, 0.37 ± 0.05 mN/mm/mm, 0.78 ± 0.08 mN/mm, and 0.94 ± 0.17 mN/mm, respectively. NAD values did not differ significantly. However, AD values for T E (1.01 ± 0.18 mN/mm), T D (1.09 ± 0.21 mN/mm), and T MAX (1.37 ± 0.24 mN/mm) were greater than ND and NAD corneas (P < 0.05). S E did not differ significantly between groups. These findings provide proof of the concept that chronic hyperglycemia from diabetes mellitus results in a phenotypically more adhesive interface between Descemet membrane and the posterior stroma in donor corneal tissue. Results of this study provide a foundation for further investigations into the impact of diabetes on the posterior cornea, eye banking, and keratoplasty. Copyright © 2016 Elsevier Ltd. All rights reserved.

Experience of Comamonas Acidovorans Keratitis with Delayed Onset and Treatment Response in Immunocompromised Cornea

PubMed Central

Lee, Sang Mok; Lee, Jae Lim; Wee, Won Ryang; Lee, Jin Hak

2008-01-01

Purpose To report 2 cases of Comamonas Acidovorans keratitis in immunocompromised cornea. Methods A complete review of the medical records of the two cases of Comamonas acidovorans keratitis. Results We found some similarities in clinical courses of two cases. Both of them showed development of keratitis during the management with corticosteroids, delayed onset, slow response to antibiotics, and relatively less affected corneal epithelium. Conclusions Comamonas Acidovorans is known as a less virulent organism. However it can cause an indolent infection that responds slowly even to adequate antibiotics therapy in immunocompromised corneas. PMID:18323706

In vitro confocal imaging of the rabbit cornea.

PubMed

Masters, B R; Paddock, S

1990-05-01

We were able to observe in vitro the fine structure of the rabbit cornea using a laser scanning confocal microscope, especially in the regions between Descemet's membrane and the epithelial basal lamina. We observed submicrometre filaments throughout the stroma with high concentrations adjacent to Descemet's membrane, and found extensive interconnecting processes between stromal keratocytes. There are numerous regions containing nerve plexuses in the stroma. We found a deeply convoluted basal lamina adjacent to the epithelium, and observed regions containing junctions between endothelial cells in fluorescent images of rabbit corneas stained with the actin-specific compound fluorescein phalloidin.

Computer driven optical keratometer and method of evaluating the shape of the cornea

NASA Technical Reports Server (NTRS)

Baroth, Edmund C. (Inventor); Mouneimme, Samih A. (Inventor)

1994-01-01

An apparatus and method for measuring the shape of the cornea utilize only one reticle to generate a pattern of rings projected onto the surface of a subject's eye. The reflected pattern is focused onto an imaging device such as a video camera and a computer compares the reflected pattern with a reference pattern stored in the computer's memory. The differences between the reflected and stored patterns are used to calculate the deformation of the cornea which may be useful for pre-and post-operative evaluation of the eye by surgeons.

[Cornea imagery and keratitis caused by processionary caterpillar hairs].

PubMed

Fournier, I; Saleh, M; Beynat, J; Creuzot-Garcher, C; Bourcier, T; Speeg-Schatz, C

2011-03-01

With their ability to migrate into the cornea and release toxins, caterpillar hairs can induce different clinical presentations such as conjunctivitis, keratoconjunctivitis, uveitis, and less frequently vitreoretinal inflammation (hyalitis, papillitis, macular edema). We report a case that occurred in Alsace (France) in a 13-years-old boy presenting with keratitis caused by caterpillar hairs. We localized them in the cornea, for the first time, using confocal microscopy and anterior segment spectral optical coherence tomography. Confocal microscopy and spectral optical coherence tomography can be useful for diagnosis and follow-up of this disease. Copyright © 2010 Elsevier Masson SAS. All rights reserved.

Sustained release and permeation of timolol from surface-modified solid lipid nanoparticles through bioengineered human cornea.

PubMed

Attama, A A; Reichl, S; Müller-Goymann, C C

2009-08-01

The aim of the study was to formulate and evaluate surface-modified solid lipid nanoparticles sustained delivery system of timolol hydrogen maleate, a prototype ocular drug using a human cornea construct. Surface-modified solid lipid nanoparticles containing timolol with and without phospholipid were formulated by melt emulsification with high-pressure homogenization and characterized by particle size, wide-angle X-ray diffraction, encapsulation efficiency, and in vitro drug release. Drug transport studies through cornea bioengineered from human donor cornea cells were carried out using a modified Franz diffusion cell and drug concentration analyzed by high-performance liquid chromatography. Results show that surface-modified solid lipid nanoparticles possessed very small particles (42.9 +/- 0.3 nm, 47.2 +/- 0.3 nm, 42.7 +/- 0.7 nm, and 37.7 +/- 0.3 nm, respectively for SM-SLN 1, SM-SLN 2, SM-SLN 3, and SM-SLN 4) with low polydispersity indices, increased encapsulation efficiency (> 44%), and sustained in vitro release compared with unmodified lipid nanoparticles whose particles were greater than 160 nm. Permeation of timolol hydrogen maleate from the surface-modified lipid nanoparticles across the cornea construct was sustained compared with timolol hydrogen maleate solution in distilled water. Surface-modified solid lipid nanoparticles could provide an efficient way of improving ocular bioavailability of timolol hydrogen maleate.

Wavelength-dependent ultraviolet induction of cyclobutane pyrimidine dimers in the human cornea.

PubMed

Mallet, Justin D; Rochette, Patrick J

2013-08-01

Exposition to ultraviolet (UV) light is involved in the initiation and the progression of skin cancer. The genotoxicity of UV light is mainly attributed to the induction of cyclobutane pyrimidine dimers (CPDs), the most abundant DNA damage generated by all UV types (UVA, B and C). The human cornea is also exposed to the harmful UV radiations, but no UV-related neoplasm has been reported in this ocular structure. The probability that a specific DNA damage leads to a mutation and eventually to cellular transformation is influenced by its formation frequency. To shed light on the genotoxic effect of sunlight in the human eye, we have analyzed CPD induction in the cornea and the iris following irradiation of ex vivo human eyes with UVA, B or C. The extent of CPD induction was used to establish the penetrance of the different UV types in the human cornea. We show that UVB- and UVC-induced CPDs are concentrated in the corneal epithelium and do not penetrate deeply beyond this corneal layer. On the other hand, UVA wavelengths penetrate deeper and induce CPDs in the entire cornea and in the first layers of the iris. Taken together, our results are undoubtedly an important step towards better understanding the consequences of UV exposure to the human eye.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cenedella, R.J.; Fleschner, C.R.

The origin of the cholesterol needed by the cornea for growth and cell turnover was addressed by comparing absolute rates of sterol synthesis with rates of sterol accumulation during early development of the rabbit. Linearity of incorporation of {sup 3}H{sub 2}O and ({sup 14}C)mevalonate into digitonin-precipitable sterols with time of incubation in vitro and a lack of accumulation of {sup 14}C in intermediates of sterol biosynthesis indicated that tritiated water can validly be used to measure rates of sterol synthesis by the cornea. The rate of sterol synthesis per unit weight of rabbit cornea was constant between 14 and 60more » days of age at an average 1.03 nmol of {sup 3}H of {sup 3}H{sub 2}O incorporated/mg dry cornea per 8 h. Essentially all of the synthesized cholesterol and most of the cholesterol mass was present in corneal epithelium. The cumulative sterol synthesized over the 46-day period studied exceeded the observed rate of cholesterol accumulation by sixfold. Cholesterol synthesized in excess of the growth requirement was likely used to support turnover of the epithelium which was estimated at 9 days. Removal of cholesterol from the cornea by excretion into tear fluid and clearance by high density lipoproteins are also considered.« less

Corneal thickness values before and after oxybuprocaine 0.4% eye drops.

PubMed

Asensio, Isabel; Rahhal, Saleh M; Alonso, Luis; Palanca-Sanfrancisco, José M; Sanchis-Gimeno, Juan A

2003-08-01

To determine changes in corneal thickness after topical anesthesia. Corneal thickness was measured before and 3 minutes after administration of two drops of oxybuprocaine 0.4% to 26 patients (26 eyes). We analyzed the corneal thickness of a control group, which was made up of 26 patients (26 eyes) before and 3 minutes after administration of two drops of saline solution. Corneal thickness was measured with the Orbscan Topography System II (Bausch Lomb Surg., Barcelona). Variations higher than +/- 10 microm were found following the instillation of 2 oxybuprocaine eye drops in eight eyes (30.76%) at the inferonasal cornea, in six eyes (23.08%) at the superotemporal, temporal and inferotemporal cornea, in five eyes (19.23%) at the nasal cornea, in three eyes (11.53%) at the central cornea, and in two eyes (7.69%) at the superonasal cornea. Nevertheless, no significant differences in the mean corneal thickness at each corneal location between the first and the second corneal thickness measurements were found in anesthetized eyes. Some individuals can present important increases and decreases in corneal thickness values after anesthetic eye drops. This effect of anesthetic eye drops must be considered by refractive surgeons when carrying out preoperative laser in situ keratomileusis corneal thickness measurements.

High-resolution, label-free two-photon imaging of diseased human corneas

NASA Astrophysics Data System (ADS)

Batista, Ana; Breunig, Hans Georg; König, Aisada; Schindele, Andreas; Hager, Tobias; Seitz, Berthold; König, Karsten

2018-03-01

The diagnosis of corneal diseases may be improved by monitoring the metabolism of cells and the structural organization of the stroma using two-photon imaging (TPI). We used TPI to assess the differences between nonpathological (NP) human corneas and corneas diagnosed with either keratoconus, Acanthamoeba keratitis, or stromal corneal scars. Images were acquired using a custom-built five-dimensional laser-scanning microscope with a broadband sub-15 femtosecond near-infrared pulsed excitation laser and a 16-channel photomultiplier tube detector in combination with a time-correlated single photon counting module. Morphological alterations of epithelial cells were observed for all pathologies. Moreover, diseased corneas showed alterations to the cells' metabolism that were revealed using the NAD(P)H free to protein-bound ratios. The mean autofluorescence lifetime of the stroma and the organization of the collagen fibers were also significantly altered due to the pathologies. We demonstrate that TPI can be used to distinguish between NP and diseased human corneas, based not only on alterations of the cells' morphology, which can also be evaluated using current clinical devices, but on additional morphological and functional features such as the organization of the stroma and the cells' metabolism. Therefore, TPI could become an efficient tool for diagnosing corneal diseases and better understanding the biological processes of the diseases.

Influence of the corneal optical zone on the point-spread function of the human eye

NASA Astrophysics Data System (ADS)

Rol, Pascal O.; Parel, Jean-Marie A.

1992-08-01

In refractive surgery, a number of surgical techniques have been developed to correct ametropia (refractive defaults) of the eye by changing the exterior shape of the cornea. Because the air-cornea interface makes up for about two thirds of the refractive power of the eye, a refractive correction can be obtained by a suitable reshaping of the cornea. Postoperatively, it is usually observed that the corneal region consists of two or more zones which are characterized by different optical parameters exhibiting in particular different focal distances. Under normal circumstances, only the central area of the cornea is involved in the formation of the retinal image. However, if part of the light entering the eye through peripheral portions of the cornea with refractive properties different from the central area can pass the pupil, an out-of-focus `ghost' image may be overlaid on the retina causing a blur. In such a case the resolution, and the contrast performance of the eye which is expected from a successful operation, may be reduced. This study is an attempt to quantify the vision blur as a function of the diameter of the central zone, i.e., the optical zone which is of importance for vision.

Corneal Wound Healing Requires IKB kinase β Signaling in Keratocytes

PubMed Central

Chen, Liang; Mongan, Maureen; Meng, Qinghang; Wang, Qin; Kao, Winston; Xia, Ying

2016-01-01

IkB kinase β (IKKβ) is a key signaling kinase for inflammatory responses, but it also plays diverse cell type-specific roles that are not yet fully understood. Here we investigated the role of IKKβ in the cornea using IkkβΔCS mice in which the Ikkβ gene was specifically deleted in the corneal stromal keratocytes. The IkkβΔCS corneas had normal morphology, transparency and thickness; however, they did not heal well from mild alkali burn injury. In contrast to the IkkβF/F corneas that restored transparency in 2 weeks after injury, over 50% of the IkkβΔCS corneas failed to fully recover. They instead developed recurrent haze with increased stromal thickness, severe inflammation and apoptosis. This pathogenesis correlated with sustained myofibroblast transformation with increased α smooth muscle actin (α-SMA) expression, higher levels of senescence β-Gal activity and scar tissue formation at the late stage of wound healing. In addition, the IkkβΔCS corneas displayed elevated expression of hemo-oxygenase-1 (HO-1), a marker of oxidative stress, and activation of stress signaling pathways with increased JNK, c-Jun and SMAD2/3 phosphorylation. These data suggest that IKKβ in keratocytes is required to repress oxidative stress and attenuate fibrogenesis and senescence in corneal wound healing. PMID:26987064

Modeling of ultrashort pulsed laser irradiation in the cornea based on parabolic and hyperbolic heat equations using electrical analogy

NASA Astrophysics Data System (ADS)

Gheitaghy, A. M.; Takabi, B.; Alizadeh, M.

2014-03-01

Hyperbolic and parabolic heat equations are formulated to study a nonperfused homogeneous transparent cornea irradiated by high power and ultrashort pulsed laser in the Laser Thermo Keratoplasty (LTK) surgery. Energy absorption inside the cornea is modeled using the Beer-Lambert law that is incorporated as an exponentially decaying heat source. The hyperbolic and parabolic bioheat models of the tissue were solved by exploiting the mathematical analogy between thermal and electrical systems, by using robust circuit simulation program called Hspice to get the solutions of simultaneous RLC and RC transmission line networks. This method can be used to rapidly calculate the temperature in laser-irradiated tissue at time and space domain. It is found that internal energy gained from the irradiated field results in a rapid rise of temperature in the cornea surface during the early heating period, while the hyperbolic wave model predicts a higher temperature rise than the classical heat diffusion model. In addition, this paper investigates and examines the effect of some critical parameters such as relaxation time, convection coefficient, radiation, tear evaporation and variable thermal conductivity of cornea. Accordingly, it is found that a better accordance between hyperbolic and parabolic models will be achieved by time.

Comparison of Cornea Module and DermaInspect for noninvasive imaging of ocular surface pathologies

NASA Astrophysics Data System (ADS)

Steven, Philipp; Müller, Maya; Koop, Norbert; Rose, Christian; Hüttmann, Gereon

2009-11-01

Minimally invasive imaging of ocular surface pathologies aims at securing clinical diagnosis without actual tissue probing. For this matter, confocal microscopy (Cornea Module) is in daily use in ophthalmic practice. Multiphoton microscopy is a new optical technique that enables high-resolution imaging and functional analysis of living tissues based on tissue autofluorescence. This study was set up to compare the potential of a multiphoton microscope (DermaInspect) to the Cornea Module. Ocular surface pathologies such as pterygia, papillomae, and nevi were investigated in vivo using the Cornea Module and imaged immediately after excision by DermaInspect. Two excitation wavelengths, fluorescence lifetime imaging and second-harmonic generation (SHG), were used to discriminate different tissue structures. Images were compared with the histopathological assessment of the samples. At wavelengths of 730 nm, multiphoton microscopy exclusively revealed cellular structures. Collagen fibrils were specifically demonstrated by second-harmonic generation. Measurements of fluorescent lifetimes enabled the highly specific detection of goblet cells, erythrocytes, and nevus-cell clusters. At the settings used, DermaInspect reaches higher resolutions than the Cornea Module and obtains additional structural information. The parallel detection of multiphoton excited autofluorescence and confocal imaging could expand the possibilities of minimally invasive investigation of the ocular surface toward functional analysis at higher resolutions.

Novel micelle carriers for cyclosporin A topical ocular delivery: in vivo cornea penetration, ocular distribution and efficacy studies.

PubMed

Di Tommaso, Claudia; Bourges, Jean-Louis; Valamanesh, Fatemeh; Trubitsyn, Gregory; Torriglia, Alicia; Jeanny, Jean-Claude; Behar-Cohen, Francine; Gurny, Robert; Möller, Michael

2012-06-01

Cornea transplantation is one of the most performed graft procedures worldwide with an impressive success rate of 90%. However, for "high-risk" patients with particular ocular diseases in addition to the required surgery, the success rate is drastically reduced to 50%. In these cases, cyclosporin A (CsA) is frequently used to prevent the cornea rejection by a systemic treatment with possible systemic side effects for the patients. To overcome these problems, it is a challenge to prepare well-tolerated topical CsA formulations. Normally high amounts of oils or surfactants are needed for the solubilization of the very hydrophobic CsA. Furthermore, it is in general difficult to obtain ocular therapeutic drug levels with topical instillations due to the corneal barriers that efficiently protect the intraocular structures from foreign substances thus also from drugs. The aim of this study was to investigate in vivo the effects of a novel CsA topical aqueous formulation. This formulation was based on nanosized polymeric micelles as drug carriers. An established rat model for the prevention of cornea graft rejection after a keratoplasty procedure was used. After instillation of the novel formulation with fluorescent labeled micelles, confocal analysis of flat-mounted corneas clearly showed that the nanosized carriers were able to penetrate into all corneal layers. The efficacy of a 0.5% CsA micelle formulation was tested and compared to a physiological saline solution and to a systemic administration of CsA. In our studies, the topical CsA treatment was carried out for 14 days, and the three parameters (a) cornea transparency, (b) edema, and (c) neovascularization were evaluated by clinical observation and scoring. Compared to the control group, the treated group showed a significant higher cornea transparency and significant lower edema after 7 and 13 days of the surgery. At the end point of the study, the neovascularization was reduced by 50% in the CsA-micelle treated animals. The success rate of cornea graft transplantation was 73% in treated animals against 25% for the control group. This result was as good as observed for a systemic CsA treatment in the same animal model. This new formulation has the same efficacy like a systemic treatment but without the serious CsA systemic side effects. Ocular drug levels of transplanted and healthy rat eyes were dosed by UPLC/MS and showed a high CsA value in the cornea (11710 ± 7530 ng(CsA)/g(tissue) and 6470 ± 1730 ng(CsA)/g(tissue), respectively). In conclusion, the applied formulation has the capacity to overcome the ocular surface barriers, the micelles formed a drug reservoir in the cornea from, where a sustained release of CsA can take place. This novel formulation for topical application of CsA is clearly an effective and well-tolerated alternative to the systemic treatment for the prevention of corneal graft rejection. Copyright © 2012 Elsevier B.V. All rights reserved.

Radius of Curvature of the Cornea--An Experiment for the Life-Science Physics Lab

ERIC Educational Resources Information Center

MacLatchy, C. S.

1978-01-01

Presents a quantitative laboratory experiment in geometrical optics. It involves the student in the measurement of the radius of curvature of the cornea and is based on an old method devised by Kohlrausch in 1839. (Author/GA)

Serial biomechanical comparison of edematous, normal, and collagen crosslinked human donor corneas using optical coherence elastography

PubMed Central

Ford, Matthew R.; Roy, Abhijit Sinha; Rollins, Andrew M.; Dupps, William J.

2014-01-01

PURPOSE To noninvasively evaluate the effects of corneal hydration and collagen crosslinking (CXL) on the mechanical behavior of the cornea. SETTING Cleveland Clinic Cole Eye Institute, Cleveland, Ohio, USA. DESIGN Experimental study. METHODS An optical coherence elastography (OCE) technique was used to measure the displacement behavior of 5 pairs of debrided human donor globes in 3 serial states as follows: edematous, normal thickness, and after riboflavin–ultraviolet-A–mediated CXL. During micromotor-controlled axial displacements with a curved goniolens at physiologic intraocular pressure (IOP), serial optical coherence tomography scans were obtained to allow high-resolution intrastromal speckle tracking and displacement measurements over the central 4.0 mm of the cornea. RESULTS With no imposed increase in IOP, the mean lateral to imposed axial displacement ratios were 0.035 μm/μm ± 0.037 (SD) in edematous corneas, 0.021 ± 0.02 μm/μm in normal thickness corneas, and 0.014 ± 0.009 μm/μm in post-CXL corneas. The differences were statistically significant (P<.05, analysis of variance) and indicated a 40% increase in lateral stromal resistance with deturgescence and a further 33% mean increase in relative stiffness with CXL. CONCLUSIONS Serial perturbations of the corneal hydration state and CXL had significant effects on corneal biomechanical behavior. With an axially applied stress from a nonapplanating contact lens, displacements along the direction of the collagen lamellae were 2 orders of magnitude lower than axial deformations. These experiments show the ability of OCE to quantify clinically relevant mechanical property differences under physiologic conditions. Financial Disclosures Proprietary or commercial disclosures are listed after the references. PMID:24767794

Experimental study of viscoelastic in the prevention of corneal endothelial desiccation injury from vitreal fluid-air exchange.

PubMed

Cekiç, Osman; Ohji, Masahito; Zheng, Yuping; Hayashi, Atsushi; Kusaka, Shunji; Tano, Yasuo

2003-05-01

To evaluate the usefulness of viscoelastic in protecting the corneal endothelium from desiccation injury associated with fluid-air exchange in a rabbit model. Experimental study. Rabbit eyes undergoing pars plana lensectomy and vitrectomy were insufflated with either dry or humidified air for 20 minutes following introduction of either Opegan (sodium hyaluronate 1.0%; Santen, Osaka, Japan) or Viscoat (sodium hyaluronate 3%-chondroitin sulfate 4%; Alcon, Tokyo, Japan) into the anterior chamber. In two other groups of rabbit eyes, the same procedure was performed without using any viscoelastic agent. Corneas obtained from rabbits undergoing surgery were compared with corneas obtained from rabbits not undergoing surgery. Potential alterations in the corneal endothelium were investigated by scanning electron microscopy, by Phalloidin-FITC staining of actin and by in vitro measurements of corneal permeability for carboxyfluorescein using a diffusion chamber. Scanning electron microscopy displayed less distortion of corneal endothelium with Opegan and Viscoat compared with the dry air-only exposed corneas. Using humidified air in Opegan and Viscoat coated corneas maintained the normal actin cytoskeleton during fluid-air exchange. Paracellular leakage was much less with Opegan and Viscoat use following infusion of dry air comparing to that of dry air-only group (P =.026 and P =.041). The difference was much more striking following humidified air infusion in Opegan or Viscoat coated corneas comparing to dry air-only infused corneas (P <.002 and P <.002). Coating of rabbit corneal endothelium with Opegan or Viscoat before fluid-air exchange largely prevents dry air damage to the endothelium. Infusion of humidified air further protects corneal endothelium during fluid-air exchange in aphakic rabbit eyes.

Epithelial innervation of human cornea: a three-dimensional study using confocal laser scanning fluorescence microscopy.

PubMed

Guthoff, Rudolf F; Wienss, Holger; Hahnel, Christian; Wree, Andreas

2005-07-01

Evaluation of a new method to visualize distribution and morphology of human corneal nerves (Adelta- and C-fibers) by means of fluorescence staining, confocal laser scanning microscopy, and 3-dimensional (3D) reconstruction. Trephinates of corneas with a diagnosis of Fuchs corneal dystrophy were sliced into layers of 200 microm thickness using a Draeger microkeratome (Storz, Germany). The anterior lamella was stained with the Life/Dead-Kit (Molecular Probes Inc.), examined by the confocal laser scanning microscope "Odyssey XL," step size between 0.5 and 1 microm, and optical sections were digitally 3D-reconstructed. Immediate staining of explanted corneas by the Life/Dead-Kit gave a complete picture of the nerves in the central human cornea. Thin nerves running parallel to the Bowman layer in the subepithelial plexus perforate the Bowman layer orthogonally through tube-like structures. Passing the Bowman layer, Adelta- and C-fibers can be clearly distinguished by fiber diameter, and, while running in the basal epithelial plexus, by their spatial arrangement. Adelta-fibers run straight and parallel to the Bowman layer underneath the basal cell layer. C-fibers, after a short run parallel to the Bowman layer, send off multiple branches penetrating epithelial cell layers orthogonally, ending blindly in invaginations of the superficial cells. In contrast to C-fibers, Adelta-fibers show characteristic bulbous formations when kinking into the basal epithelial plexus. Ex-vivo fluorescence staining of the cornea and 3D reconstructions of confocal scans provide a fast and easily reproducible tool to visualize nerves of the anterior living cornea at high resolution. This may help to clarify gross variations of nerve fiber patterns under various clinical and experimental conditions.

Infrared irradiation alters the expression of matrix metalloproteinases and glycosaminoglycans in the cornea and crystalline lens.

PubMed

Dadoukis, Panagiotis; Klagas, Ioannis; Komnenou, Anastasia; Karakiulakis, George; Karoutis, Athanasios; Karampatakis, Vassilios; Papakonstantinou, Eleni

2013-08-01

Prolonged exposure to infrared (IR) radiation is associated with different types of damage to cornea and lens. The aim of our study was to investigate the effect of acute and chronic exposure to IR radiation on the activity of matrix metalloproteinase-2 (MMP-2) and MMP-9 and on the expression of glycosaminoglycans (GAG) in the rabbit cornea and crystalline lens. New Zealand rabbits were subjected to IR radiation for 4 months (chronic exposure to IR) or to normal light (control group). In experiments regarding acute exposure, animals were subjected to IR radiation or normal light for 12 h, in the presence of 0.1% diclofenac sodium (eye drops instilled in the right eye of animals) or saline (instilled in the left eye of animals). The cornea and lens were dissected away and homogenized. The activity of MMP-2 and MMP-9 was assayed by gelatine zymography. Total GAG were isolated from tissue specimens after lipid extraction and extensive digestion with pronase and DNase and characterized by treatment with GAG-degrading enzymes, followed by electrophoresis on cellulose acetate membranes. Acute or chronic exposure to IR radiation induced the activity of MMP-2 in cornea and lens, whereas only acute IR radiation increased the content of heparan sulphate in crystalline lens. Local administration of diclofenac sodium did not prevent the above effects of acute IR radiation. The detrimental effects of excessive or prolonged exposure of the eyes to IR radiation are associated with induced activity of MMP-2 in cornea and lens and alterations in the content of heparan sulphate in lens. Thus, MMP and GAG may offer alternative targets for pharmacological intervention to confront ocular damages associated with IR radiation.

Customized Corneal Cross-Linking-A Mathematical Model.

PubMed

Caruso, Ciro; Epstein, Robert L; Ostacolo, Carmine; Pacente, Luigi; Troisi, Salvatore; Barbaro, Gaetano

2017-05-01

To improve the safety, reproducibility, and depth of effect of corneal cross-linking with the ultraviolet A (UV-A) exposure time and fluence customized according to the corneal thickness. Twelve human corneas were used for the experimental protocol. They were soaked using a transepithelial (EPI-ON) technique using riboflavin with the permeation enhancer vitamin E-tocopheryl polyethylene glycol succinate. The corneas were then placed on microscope slides and irradiated at 3 mW/cm for 30 minutes. The UV-A output parameters were measured to build a new equation describing the time-dependent loss of endothelial protection induced by riboflavin during cross-linking, as well as a pachymetry-dependent and exposure time-dependent prescription for input UV-A fluence. The proposed equation was used to establish graphs prescribing the maximum UV-A fluence input versus exposure time that always maintains corneal endothelium exposure below toxicity limits. Analysis modifying the Lambert-Beer law for riboflavin oxidation leads to graphs of the maximum safe level of UV-A radiation fluence versus the time applied and thickness of the treated cornea. These graphs prescribe UV-A fluence levels below 1.8 mW/cm for corneas of thickness 540 μm down to 1.2 mW/cm for corneas of thickness 350 μm. Irradiation times are typically below 15 minutes. The experimental and mathematical analyses establish the basis for graphs that prescribe maximum safe fluence and UV-A exposure time for corneas of different thicknesses. Because this clinically tested protocol specifies a corneal surface clear of shielding riboflavin on the corneal surface during UV-A irradiation, it allows for shorter UV-A irradiation time and lower fluence than in the Dresden protocol.

Cornea and ocular lens visualized with three-dimensional confocal microscopy

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1992-08-01

This paper demonstrates the advantages of three-dimensional reconstruction of the cornea and the ocular crystalline lens by confocal microscopy and volume rendering computer techniques. The advantages of noninvasive observation of ocular structures in living, unstained, unfixed tissue include the following: the tissue is in a natural living state without the artifacts of fixation, mechanical sectioning, and staining; the three-dimensional structure can be observed from any view point and quantitatively analyzed; the dynamics of morphological changes can be studied; and the use of confocal microscopic observation results in a reduction of the number of animals required for ocular morphometric studies. The main advantage is that the dynamic morphology of ocular structures can be investigated in living ocular tissue. A laser scanning confocal microscope was used in the reflected light mode to obtain the two- dimensional images from the cornea and the ocular lens of a freshly enucleated rabbit eye. The light source was an argon ion laser with 488 nm wavelength. The microscope objective was a Leitz 25X, NA 0.6 water immersion lens. The 400 micron thick cornea was optically sectioned into 133, three micron sections. The semi-transparent cornea and the in-situ ocular lens was visualized as high resolution, high contrast two-dimensional images. The under sampling resulted in a three-dimensional visualization rendering in which the corneal thickness (z-axis) is compressed. The structures observed in the cornea include: superficial epithelial cells and their nuclei, basal epithelial cells and their `beaded' cell borders, basal lamina, nerve plexus, nerve fibers, free nerve endings in the basal epithelial cells, nuclei of stromal keratocytes, and endothelial cells. The structures observed in the in-situ ocular lens include: lens capsule, lens epithelial cells, and individual lens fibers.

Corneal surface glycosylation is modulated by IL-1R and Pseudomonas aeruginosa challenge but is insufficient for inhibiting bacterial binding.

PubMed

Jolly, Amber L; Agarwal, Paresh; Metruccio, Matteo M E; Spiciarich, David R; Evans, David J; Bertozzi, Carolyn R; Fleiszig, Suzanne M J

2017-06-01

Cell surface glycosylation is thought to be involved in barrier function against microbes at mucosal surfaces. Previously we showed that the epithelium of healthy mouse corneas becomes vulnerable to Pseudomonas aeruginosa adhesion if it lacks the innate defense protein MyD88 (myeloid differentiation primary response gene 88), or after superficial injury by blotting with tissue paper. Here we explored their effect on corneal surface glycosylation using a metabolic label, tetra-acetylated N -azidoacetylgalactosamine (Ac 4 GalNAz). Ac 4 GalNAz treatment labeled the surface of healthy mouse corneas, leaving most cells viable, and bacteria preferentially associated with GalNAz-labeled regions. Surprisingly, corneas from MyD88 -/- mice displayed similar GalNAz labeling to wild-type corneas, but labeling was reduced and patchy on IL-1 receptor (IL-1R)-knockout mouse corneas ( P < 0.05, ANOVA). Tissue paper blotting removed GalNAz-labeled surface cells, causing DAPI labeling (permeabilization) of underlying cells. MS of material collected on the tissue paper blots revealed 67 GalNAz-labeled proteins, including intracellular proteins. These data show that the normal distribution of surface glycosylation requires IL-1R, but not MyD88, and is not sufficient to prevent bacterial binding. They also suggest increased P. aeruginosa adhesion to MyD88 -/- and blotted corneas is not due to reduction in total surface glycosylation, and for tissue paper blotting is likely due to cell permeabilization.-Jolly, A. L., Agarwal, P., Metruccio, M. M. E., Spiciarich, D. R., Evans, D. J., Bertozzi, C. R., Fleiszig, S. M. J. Corneal surface glycosylation is modulated by IL-1R and Pseudomonas aeruginosa challenge but is insufficient for inhibiting bacterial binding. © FASEB.

Lens and cornea lesions of rats fed corn syrup and the protective effects of alpha lipoic acid.

PubMed

Gunes, Alime; Ozmen, Ozlem; Saygın, Mustafa; Ascı, Halil; Tok, Levent; Tok, Ozlem; Dıncoglu, Dılnur

2016-03-01

To examine the pathological findings that occurred in the lens and cornea and biochemical findings in the lens of rats fed with corn syrup and the protective effects of alpha lipoic acid (ALA). Twenty-four rats were randomly divided into three groups. Group I served as the control group. Group II was used as the study group; the rats were treated with 30% corn sugar solution for 10 weeks. Group III was the treatment group. Corn syrup was given by the oral route to the rats during the study, and ALA (100 mg/kg) was added to the treatment 4 weeks after the study began. At the end of the experiment, central corneal thickness (CCT) was measured in all rats with an ultrasonic pachymeter. Then the right eyes of the rats were enucleated for histopathological examination of the cornea and lens. The left lenses were homogenized for biochemical analyses. The lenses of the rats treated with corn syrup revealed severe damage; many lens fibers appeared swollen and ruptured with large vacuoles near the lens epithelium. In addition, malondialdehyde (MDA) levels, a parameter of oxidative stress, increased but not significantly in Group II; however. ALA treatment decreased MDA levels significantly. Antioxidant enzyme and catalase (CAT) activities were significantly decreased in Group II, and ALA treatment increased these activities; however, the increase was not significant. Changes were observed in the cornea such as epithelial alterations, subepithelial vacuolizations, collagen fibers loss in the stromal layer, interruptions in the subepithelial basement membrane and central corneal thickening. Corn syrup can cause severe damage in rat lenses and corneas. However, ALA ameliorates the effect of corn syrup-related lesions on the cornea and lens.

Thermodynamic measurement after cooling the cornea with intact epithelium and lid manipulation.

PubMed

de Ortueta, Diego; Magnago, Thomas; Arba-Mosquera, Samuel

2015-01-01

To characterize the rate of change of ocular surface temperature (OST) under lid manipulation after cooling the intact cornea with balanced salt solution (BSS). In a patient for refractive surgery, prior to the ablation, the temperature of the cornea was continuously recorded with a high speed infrared (350Hz) camera. Two millilitre of chilled BSS with a temperature of 8.6°Celsius (°C) was instilled for about 3s. Using exponential functions, the three contributions have been determined, subjacent corneal layers, environment, and chilled BSS. The mean temperature of the cornea preoperatively was 34.5°C. After applying the chilled BSS the temperature decreased about 14°C down to an OST of 20°C and the time needed afterwards to get the normal (OST) temperature of about 30°C was 40s. Due to the inserted speculum and missing blink, OST did not reach the original OST of 34.5°C and faded at about 32.5°C. According to our best fitted model, absolute value of each contributing component was 31.4°C (subjacent corneal layers), 26.8°C (environment) and 8.6°C (BSS). Applying chilled BSS to the cornea quickly reduces the temperature of the cornea with a thermal relaxation time of 3s and a amplitude decrease of 8.6°C. This together with a relaxation time of 7s for subjacent corneal layers, and 184s for environment after instillation of BSS combined with a well-controlled environment provides a period of 40s of corneal temperature below baseline, which may be of clinical benefit when applying chilled BSS immediately before or immediately after ablation. Copyright © 2014 Spanish General Council of Optometry. Published by Elsevier Espana. All rights reserved.

Analysis of the Viscoelastic Properties of the Human Cornea Using Scheimpflug Imaging in Inflation Experiment of Eye Globes

PubMed Central

Lombardo, Giuseppe; Serrao, Sebastiano; Rosati, Marianna; Lombardo, Marco

2014-01-01

Purpose To demonstrate a Scheimpflug-based imaging procedure for investigating the depth- and time-dependent strain response of the human cornea to inflation testing of whole eye globes. Methods Six specimens, three of which with intact corneal epithelium, were mounted in a customized apparatus within a humidity and temperature-monitored wet chamber. Each specimen was subjected to two mechanical tests in order to measure corneal strain resulting from application of cyclic (cyclic regimen) and constant (creep regimen) stress by changing the intra-ocular pressure (IOP) within physiological ranges (18–42 mmHg). Corneal shape changes were analyzed as a function of IOP and both corneal stress-strain curves and creep curves were generated. Results The procedure was highly accurate and repeatable. Upon cyclic stress application, a biomechanical corneal elasticity gradient was found in the front-back direction. The average Young's modulus of the anterior cornea ranged between 2.28±0.87 MPa and 3.30±0.90 MPa in specimens with and without intact epithelium (P = 0.05) respectively. The Young's modulus of the posterior cornea was on average 0.21±0.09 MPa and 0.17±0.06 MPa (P>0.05) respectively. The time-dependent strain response of the cornea to creep testing was quantified by fitting data to a modified Zener model for extracting both the relaxation time and compliance function. Conclusion Cyclic and creep mechanical tests are valuable for investigating the strain response of the intact human cornea within physiological IOP ranges, providing meaningful results that can be translated to clinic. The presence of epithelium influences the results of anterior corneal shape changes when monitoring deformation via Scheimpflug imaging in inflation experiments of whole eye globes. PMID:25397674

Assessment of UVA-Riboflavin Corneal Cross-Linking Using Small Amplitude Oscillatory Shear Measurements.

PubMed

Aslanides, Ioannis M; Dessi, Claudia; Georgoudis, Panagiotis; Charalambidis, Georgios; Vlassopoulos, Dimitris; Coutsolelos, Athanassios G; Kymionis, George; Mukherjee, Achyut; Kitsopoulos, Theofanis N

2016-04-01

The effect of ultraviolet (UV)-riboflavin cross-linking (CXL) has been measured primarily using the strip extensometry technique. We propose a simple and reliable methodology for the assessment of CXL treatment by using an established rheologic protocol based on small amplitude oscillatory shear (SAOS) measurements. It provides information on the average cross-link density and the elastic modulus of treated cornea samples. Three fresh postmortem porcine corneas were used to study the feasibility of the technique, one serving as control and two receiving corneal collagen cross-linking treatment. Subsequently, five pairs of fresh postmortem porcine corneas received corneal collagen cross-linking treatment with riboflavin and UVA-irradiation (370 nm; irradiance of 3 mW/cm2) for 30 minutes (Dresden protocol); the contralateral porcine corneas were used as control samples. After the treatment, the linear viscoelastic moduli of the corneal samples were measured using SAOS measurements and the average cross-linking densities extracted. For all cases investigated, the dynamic moduli of the cross-linked corneas were higher compared to those of the corresponding control samples. The increase of the elastic modulus of the treated samples was between 122% and 1750%. The difference was statistically significant for all tested samples (P = 0.018, 2-tailed t-test). We report a simple and accurate methodology for quantifying the effects of cross-linking on porcine corneas treated with the Dresden protocol by means of SAOS measurements in the linear regime. The measured dynamic moduli, elastic and viscous modulus, represent the energy storage and energy dissipation, respectively. Hence, they provide a means to assess the changing physical properties of the cross-linked collagen networks after CXL treatment.

[Significance of laser confocal tomography in diagnosis and monitoring of keratoconjunctivitis sicca].

PubMed

Safonova, T N; Gladkova, О V; Boev, V I

2016-01-01

Laser confocal tomography of the cornea enables studying ultrathin sections of corneal layers, which provides additional reliable information on tissue changes in keratoconjunctivitis sicca (KCS). To assess the significance of laser confocal tomography of the cornea in the diagnosis and monitoring of KCS. We investigated 38 eyes of 30 patients with severe KCS. The patients were divided into two groups. Group 1 (15 patients, 19 eyes) was prescribed cyclosporine А 0.05% instillations 2 times daily, artificial tears, and soft contact lenses. Group 2 (15 patients, 19 eyes) received only instillations of cyclosporine А 0.05% 2 times daily and artificial tears. Besides standard ophthalmic examination, additional tests were performed, namely Schirmer's test, tear break-up time test, fluorescein eye stain test, tear osmolarity test (TearLab System, USA), and Heidelberg retinal tomography of the cornea (HRT, Heidelberg Engineering GmbH, Germany). HRT findings revealed a 3 times shorter epithelization period and faster recovery of corneal transparency in group 1 as compared to group 2 (1.5 and 4.5 months, respectively). There was also an evident reduction in the number of immune cells in the cornea, most pronounced in group 1 at 3 months, which is indicative of inflammation termination. The use of HRT of the cornea in KCS patients allows real-time cellular level observation of corneal changes, which together with clinical findings and diagnostic tests not only confirms the diagnosis but also determines treatment effectiveness. It has been also found that soft contact lenses accelerate epithelization of the cornea and relieve inflammation of the ocular surface in KCS patients under cyclosporine A 0.05% instillation therapy. Transparency of financial activity: the authors have no financial interest in the submitted materials and methods.

Safety profile of topical VEGF neutralization at the cornea.

PubMed

Bock, Felix; Onderka, Jasmine; Rummelt, Carmen; Dietrich, Tina; Bachmann, Björn; Kruse, Friedrich E; Schlötzer-Schrehardt, Ursula; Cursiefen, Claus

2009-05-01

Bevacizumab eyedrops inhibit corneal neovascularization. The purpose of this study was to analyze the safety profile of VEGF-A neutralization at the ocular surface. Bevacizumab eyedrops (5 mg/mL) and an antimurine VEGF-A antibody (250 microg/mL) were applied to normal murine corneas five times a day for 7 and 14 days. Subsequently, corneas were analyzed for morphologic changes by light and electron microscopy. In a mouse model of corneal epithelial abrasion, the effects of topically applied anti-VEGF antibodies on epithelial wound healing were analyzed: the treatment group received bevacizumab (5 mg/mL) or the antimurine VEGF-A antibody (250 microg/mL) as eyedrops, and the control group received an equal volume of saline solution. After 12, 18, and 24 hours, corneas were photographed in vivo with and without fluorescein staining for morphometry. Afterwards the mice were killed, and eyes were removed for histology, immunohistochemistry with Ki67/DAPI, and electron microscopy. The effect of midterm anti-VEGF therapy on corneal nerve density was assessed by staining corneas treated with an FITC-conjugated anti-neurofilament antibody and morphometric analysis. Murine corneas treated with two different types of anti-VEGF antibody eyedrops did not show obvious corneal morphologic changes at the light and electron microscopic levels. Furthermore, anti-VEGF antibody eyedrops had no significant impact on the wound healing process after corneal epithelial injury or on normal murine corneal nerve fiber density. Topical neutralization of VEGF-A at the corneal surface does not have significant side effects on normal corneal epithelial wound healing, normal corneal integrity, or normal nerve fiber density. Therefore, anti-VEGF eyedrops seem to be a relatively safe option to treat corneal neovascularization.

Corneal endothelium: developmental strategies for regeneration

PubMed Central

Zavala, J; López Jaime, G R; Rodríguez Barrientos, C A; Valdez-Garcia, J

2013-01-01

The main treatment available for restoration of the corneal endothelium is keratoplasty. This procedure is faced with several difficulties, including the shortage of donor tissue, post-surgical complications associated with the use of drugs to prevent immune rejection, and a significant increase in the occurrence of glaucoma. Recently, surgical procedures such as Descemet's stripping endothelial keratoplasty have focused on the transplant of corneal endothelium, yielding better visual results but still facing the need for donor tissue. The emergent strategies in the field of cell biology and tissue cultivation of corneal endothelial cells aim at the production of transplantable endothelial cell sheets. Cell therapy focuses on the culture of corneal endothelial cells retrieved from the donor, in the donor's cornea, followed by transplantation into the recipient. Recently, research has focused on overcoming the challenge of harvesting human corneal endothelial cells and the generation of new biomembranes to be used as cell scaffolds in surgical procedures. The use of corneal endothelial precursors from the peripheral cornea has also demonstrated to be effective and represents a valuable tool for reducing the risk of rejection in allogeneic transplants. Several animal model reports also support the use of adult stem cells as therapy for corneal diseases. Current results represent important progresses in the development of new strategies based on alternative sources of tissue for the treatment of corneal endotheliopathies. Different databases were used to search literature: PubMed, Google Books, MD Consult, Google Scholar, Gene Cards, and NCBI Books. The main search terms used were: ‘cornea AND embryology AND transcription factors', ‘human endothelial keratoplasty AND risk factors', ‘(cornea OR corneal) AND (endothelium OR endothelial) AND cell culture', ‘mesenchymal stem cells AND cell therapy', ‘mesenchymal stem cells AND cornea', and ‘stem cells AND (cornea OR corneal) AND (endothelial OR endothelium)'. PMID:23470788

Noninvasive intratissue refractive index shaping (IRIS) of the cornea with blue femtosecond laser light.

PubMed

Xu, Lisen; Knox, Wayne H; DeMagistris, Margaret; Wang, Nadan; Huxlin, Krystel R

2011-10-17

To test the feasibility of intratissue refractive index shaping (IRIS) in living corneas by using 400-nm femtosecond (fs) laser pulses (blue-IRIS). To test the hypothesis that the intrinsic two-photon absorption of the cornea allows blue-IRIS to be performed with greater efficacy than when using 800-nm femtosecond laser pulses. Fresh cat corneas were obtained postmortem and cut into six wedges. Blue laser pulses at 400 nm, with 100-fs pulse duration at 80 MHz were used to micromachine phase gratings into each corneal wedge at scanning speeds from 1 to 15 mm/s. Grating lines were 1 μm wide, 5 μm apart, and 150 μm below the anterior corneal surface. Refractive index (RI) changes in micromachined regions were measured immediately by recording the diffraction efficiency of inscribed gratings. Six hours later, the corneas were processed for histology, and TUNEL staining was performed to assess whether blue-IRIS causes cell death. Scanning at 1 and 2 mm/s caused overt corneal damage in the form of bubbles and burns. At faster scanning speeds (5, 10, and 15 mm/s), phase gratings were created in the corneal stroma, which were shown to be pure RI changes ranging from 0.037 to 0.021 in magnitude. The magnitude of RI change was inversely related to scanning speed. TUNEL staining showed cell death only around bubbles and burns. Blue-IRIS can be performed safely and effectively in living cornea. Compared with near-infrared laser pulses, blue-IRIS enhances both achievable RI change and scanning speed without the need to dope the tissue with two-photon sensitizers, increasing the clinical applicability of this technique.

Noninvasive Intratissue Refractive Index Shaping (IRIS) of the Cornea with Blue Femtosecond Laser Light

PubMed Central

Xu, Lisen; Knox, Wayne H.; DeMagistris, Margaret; Wang, Nadan

2011-01-01

Purpose. To test the feasibility of intratissue refractive index shaping (IRIS) in living corneas by using 400-nm femtosecond (fs) laser pulses (blue-IRIS). To test the hypothesis that the intrinsic two-photon absorption of the cornea allows blue-IRIS to be performed with greater efficacy than when using 800-nm femtosecond laser pulses. Methods. Fresh cat corneas were obtained postmortem and cut into six wedges. Blue laser pulses at 400 nm, with 100-fs pulse duration at 80 MHz were used to micromachine phase gratings into each corneal wedge at scanning speeds from 1 to 15 mm/s. Grating lines were 1 μm wide, 5 μm apart, and 150 μm below the anterior corneal surface. Refractive index (RI) changes in micromachined regions were measured immediately by recording the diffraction efficiency of inscribed gratings. Six hours later, the corneas were processed for histology, and TUNEL staining was performed to assess whether blue-IRIS causes cell death. Results. Scanning at 1 and 2 mm/s caused overt corneal damage in the form of bubbles and burns. At faster scanning speeds (5, 10, and 15 mm/s), phase gratings were created in the corneal stroma, which were shown to be pure RI changes ranging from 0.037 to 0.021 in magnitude. The magnitude of RI change was inversely related to scanning speed. TUNEL staining showed cell death only around bubbles and burns. Conclusions. Blue-IRIS can be performed safely and effectively in living cornea. Compared with near-infrared laser pulses, blue-IRIS enhances both achievable RI change and scanning speed without the need to dope the tissue with two-photon sensitizers, increasing the clinical applicability of this technique. PMID:21931133

Genetic evidence for the coordinated regulation of collagen fibrillogenesis in the cornea by decorin and biglycan.

PubMed

Zhang, Guiyun; Chen, Shoujun; Goldoni, Silvia; Calder, Bennett W; Simpson, Holly C; Owens, Rick T; McQuillan, David J; Young, Marian F; Iozzo, Renato V; Birk, David E

2009-03-27

Decorin and biglycan are class I small leucine-rich proteoglycans (SLRPs) involved in regulation of collagen fibril and matrix assembly. We hypothesize that tissue-specific matrix assembly, such as in the cornea, requires a coordinate regulation involving multiple SLRPs. To this end, we investigated the expression of decorin and biglycan in the cornea of mice deficient in either SLRP gene and in double-mutant mice. Decorin and biglycan exhibited overlapping spatial expression patterns throughout the corneal stroma with differential temporal expression. Whereas decorin was expressed at relatively high levels in all developmental stages, biglycan expression was high early, decreased during development, and was present at very low levels in the mature cornea. Ultrastructural analyses demonstrated comparable fibril structure in the decorin- and biglycan-null corneas compared with wild-type controls. We found a compensatory up-regulation of biglycan gene expression in the decorin-deficient mice, but not the reverse. Notably, the corneas of compound decorin/biglycan-null mice showed severe disruption in fibril structure and organization, especially affecting the posterior corneal regions, corroborating the idea that biglycan compensates for the loss of decorin. Fibrillogenesis assays using recombinant decorin and biglycan confirmed a functional compensation, with both having similar effects at high SLRP/collagen ratios. However, at low ratios decorin was a more efficient regulator. The use of proteoglycan or protein core yielded comparable results. These findings provide firm genetic evidence for an interaction of decorin and biglycan during corneal development and further suggest that decorin has a primary role in regulating fibril assembly, a function that can be fine-tuned by biglycan during early development.

Ophthalmic applications of confocal microscopy: diagnostics, refractive surgery, and eye banking

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1990-11-01

Confocal microscopy of ocular tissue provides two advantages over traditional imaging techniques: increased range and transverse resolution and increased contrast. The semitransparent cornea and ocular lens in the living eye can be optically sectioned and observed by reflected light confocal microscopy. Within the cornea we observed various cell components nerve fibers nerve cell bodies and fibrous networks. The confocal microscopic images from the in-situ ocular lens show the lens capsule the lens epithelium and the individual lens fibrils. All of the reflected light confocal microscopic images have high contrast and high resolution. Some of the applications of confocal imaging in ophthalmology include: diagnostics of the cornea and the ocular lens examination prior to and after refractive surgery examination of intraocular lenses (IOL) and examination of eye bank material. Other ophthalmic uses of confocal imaging include: studies of wound healing therapeutics and the effects of contact lenses on the cornea. The proposed features of a clinical confocal microscope are reviewed. 2.

Development of cataract and corneal opacity in mice due to radon exposure

NASA Astrophysics Data System (ADS)

Abdelkawi, S. A.; Abo-Elmagd, M.; Soliman, H. A.

This work investigates the radiation damage on the eye of albino mice exposed to effective radon doses ranging from 20.92 to 83.68 mSv. These doses were taken over 2-8 weeks using a radon chamber constructed by the National Institute for Standard (Egypt). The guidance on the quality assurance program for radon measurements was followed. Therefore, the estimated doses received by the laboratory animals meet the requirements of national standardE The refractive index(RI) and protein concentration were measured for soluble proteins of both corneas and lenses. In addition, the sodium dodecyle sulfate polyacrylamide gel electrophoresis (SDSPAGE) technique was used. The results show increasing of the RI of both cornea and lens proteins and decreasing in total protein concentration of exposed animals. These results were accompanied with changes in the SDSPAGE profile for both cornea and lens. Therefore, radon exposure produces substantial hazards to the cornea and lens of experimental animals and has a crucial role in the development of cataract and corneal opacity.

The mouse cornea micropocket angiogenesis assay.

PubMed

Rogers, Michael S; Birsner, Amy E; D'Amato, Robert J

2007-01-01

The mouse corneal micropocket angiogenesis assay uses the avascular cornea as a canvas to study angiogenesis in vivo. Through the use of standardized slow-release pellets, a predictable angiogenic response is generated over the course of 5 d and then quantified. Uniform slow-release pellets are prepared by mixing purified angiogenic growth factors such as basic fibroblast growth factor or vascular endothelial growth factor with sucralfate (a stabilizer) and Hydron (poly-HEMA (poly(2-hydroxyethyl methacrylate)) to allow slow release). This mixture is applied to a mesh that controls unit size and then allowed to harden. A micropocket is surgically created in the mouse cornea and a pellet implanted. Five days later, the area of the cornea overgrown by the angiogenic response is measured using a slit lamp. A skilled investigator can implant and grade 40 eyes in about 2.5 h. The results of the assay are used to assess the ability of potential therapeutic molecules or genetic differences to modulate angiogenesis in vivo.

High-precision double-frequency interferometric measurement of the cornea shape

NASA Astrophysics Data System (ADS)

Molebny, Vasyl V.; Pallikaris, Ioannis G.; Naoumidis, Leonidas P.; Smirnov, Eugene M.; Ilchenko, Leonid M.; Goncharov, Vadym O.

1996-11-01

To measure the shape of the cornea and its declinations from the necessary values before and after PRK operation, s well as the shape of other spherical objects like artificial pupil, a technique was used of double-frequency dual-beam interferometry. The technique is based on determination of the optical path difference between two neighboring laser beams, reflected from the cornea or other surface under investigation. Knowing the distance between the beams on the investigated shape. The shape itself is reconstructed by along-line integration. To adjust the wavefront orientation of the laser beam to the spherical shape of the cornea or artificial pupil in the course of scanning, additional lens is involved. Signal-to-noise ratio is ameliorated excluding losses in the acousto-optic deflectors. Polarization selection is realized for choosing the signal needed for measurement. 2D image presentation is accompanied by convenient PC accessories, permitting precise cross-section measurements along selected directions. Sensitivity of the order of 10-2 micrometers is achieved.

Second Harmonic Generation Imaging Analysis of Collagen Arrangement in Human Cornea.

PubMed

Park, Choul Yong; Lee, Jimmy K; Chuck, Roy S

2015-08-01

To describe the horizontal arrangement of human corneal collagen bundles by using second harmonic generation (SHG) imaging. Human corneas were imaged with an inverted two photon excitation fluorescence microscope. The excitation laser (Ti:Sapphire) was tuned to 850 nm. Backscatter signals of SHG were collected through a 425/30-nm bandpass emission filter. Multiple, consecutive, and overlapping image stacks (z-stacks) were acquired to generate three dimensional data sets. ImageJ software was used to analyze the arrangement pattern (irregularity) of collagen bundles at each image plane. Collagen bundles in the corneal lamellae demonstrated a complex layout merging and splitting within a single lamellar plane. The patterns were significantly different in the superficial and limbal cornea when compared with deep and central regions. Collagen bundles were smaller in the superficial layer and larger in deep lamellae. By using SHG imaging, the horizontal arrangement of corneal collagen bundles was elucidated at different depths and focal regions of the human cornea.

Elastography methods applicable to the eye

NASA Astrophysics Data System (ADS)

Khan, Altaf A.; Cortina, Soledad M.; Chamon, Wallace; Royston, Thomas J.

2014-02-01

Elastography is the mapping of tissues and cells by their respective mechanical properties, such as elasticity and viscosity. Our interest primarily lies in the human eye. Combining Scanning Laser Doppler Vibrometry (SLDV) with geometrically focused mechanical vibratory excitations of the cornea, it is possible to reconstruct these mechanical properties of the cornea. Experiments were conducted on phantom corneas as well as excised donor human corneas to test feasibility and derive a method of modeling. Finite element analysis was used to recreate the phantom studies and corroborate with the experimental data. Results are in close agreement. To further expand the study, lamb eyes were used in MR Elastography studies. 3D wave reconstruction was created and elastography maps were obtained. With MR Elastography, it would be possible to noninvasively measure mechanical properties of anatomical features not visible to SLDV, such as the lens and retina. Future plans include creating a more robust finite element model, improving the SLDV method for in-vivo application, and continuing experiments with MR Elastography.

Noninvasive optical coherence tomography monitoring of structure and hydration changes of human corneas in different preservation media

NASA Astrophysics Data System (ADS)

Wu, Yicong; Clarke, Dominic; Mathew, Aby; Nicoud, Ian; Li, Xingde

2011-02-01

The influence of different tissue preservation (a test solution under development and a standard storage solution) on human cornea morphology, refractive index and hydration was assessed noninvasively by ultrahigh-resolution optical coherence tomography (OCT) over time. For 28 days' or 15 days' storage in the preservation media, corneas in the two media exhibited different structural changes with different onset times including epithelial desquamation, edema-induced cornea thickening and change in tissue refractive index. It was found that the variation of the group refractive index over time was only about 2%, while 25% variation of hydration was observed in the storage and subsequent return to normothermic conditions in both preservation media. The results suggest the two media involved different but correlated preservation mechanisms. This study demonstrates that the noncontact, noninvasive, and high-resolution OCT is a powerful tool for noninvasive characterization of tissue morphological changes and hydration process and for assessment of the effects of preservation media on stored tissue integrity. Engineers.

Using an ultrasound elasticity microscope to map three-dimensional strain in a porcine cornea.

PubMed

Hollman, Kyle W; Shtein, Roni M; Tripathy, Sakya; Kim, Kang

2013-08-01

An ultrasound elasticity microscope was used to map 3-D strain volume in an ex vivo porcine cornea to illustrate its ability to measure the mechanical properties of this tissue. Mechanical properties of the cornea play an important role in its function and, therefore, also in ophthalmic diseases such as kerataconus and corneal ectasia. The ultrasound elasticity microscope combines a tightly focused high-frequency transducer with confocal scanning to produce high-quality speckle over the entire volume of tissue. This system and the analysis were able to generate volume maps of compressional strain in all three directions for porcine corneal tissue, more information than any previous study has reported. Strain volume maps indicated features of the cornea and mechanical behavior as expected. These results constitute a step toward better understanding of corneal mechanics and better treatment of corneal diseases. Copyright © 2013 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

The role of corneal endothelial morphology in graft assessment and prediction of endothelial cell loss during organ culture of human donor corneas.

PubMed

Hermel, Martin; Salla, Sabine; Fuest, Matthias; Walter, Peter

2017-03-01

Endothelial assessment is crucial in the release of corneas for grafting. We retrospectively analysed the role of endothelial morphology parameters in predicting endothelial cell loss during organ culture. Human donor corneas were cultured in minimal essential medium with 2% fetal calf serum and antibiotics. Initial endothelial morphology was assessed microscopically using score parameters polymegethism (POL), pleomorphism (PLE), granulation (GRA), vacuolization (VAC), segmentation of cell membranes (SEG), Descemet's folds (DF), trypan blue-positive cells (TBPC) and endothelial cell-free areas (ECFA). Some corneas were primarily rejected based on endothelial assessment. Endothelial cell density (ECD) was assessed at the beginning (I-ECD) and end of culture. Corneas were then placed in dehydration medium (as above + 5% dextran 500). In a subgroup, ECD was reassessed after dehydration. Endothelial cell loss during culture (ECL@Culture) and culture+dehydration (ECL-Culture&Dehydration) were calculated. Data were given as mean ± SD and analysed using multiple linear and logistic regression. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. I-ECD was 2812 ± 360/mm 2 (n = 2356). The decision to reject a cornea due to endothelial assessment was associated negatively with I-ECD (OR = 0.77/100 cells, CI 0.7-0.82) and positively with ECFA (OR = 2.7, CI 1.69-4.35), SEG (OR =1.3, CI 1.01-1.68) and donor age (OR = 1.26/decade, CI 1.33-1.41). ECL@Culture was 153 ± 201/mm 2 (n = 1277), ECL@Culture&Dehydration was 169 ± 183/mm 2 (n = 918). ECL@Culture was associated positively with donor age, I-ECD, GRA and TBPC, and negatively with PLE, and DF. ECL@Culture&Dehydration was associated positively with age, sex, initial ECD, POL, PLE, VAC and TBPC. Morphological parameters displayed associations with the exclusion of corneas from culture and with endothelial cell loss. Appropriate parameter selection for screening purposes may help improve graft quality. © 2016 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

A new method of detecting changes in corneal health in response to toxic insults.

PubMed

Khan, Mohammad Faisal Jamal; Nag, Tapas C; Igathinathane, C; Osuagwu, Uchechukwu L; Rubini, Michele

2015-11-01

The size and arrangement of stromal collagen fibrils (CFs) influence the optical properties of the cornea and hence its function. The spatial arrangement of the collagen is still questionable in relation to the diameter of collagen fibril. In the present study, we introduce a new parameter, edge-fibrillar distance (EFD) to measure how two collagen fibrils are spaced with respect to their closest edges and their spatial distribution through normalized standard deviation of EFD (NSDEFD) accessed through the application of two commercially available multipurpose solutions (MPS): ReNu and Hippia. The corneal buttons were soaked separately in ReNu and Hippia MPS for five hours, fixed overnight in 2.5% glutaraldehyde containing cuprolinic blue and processed for transmission electron microscopy. The electron micrographs were processed using ImageJ user-coded plugin. Statistical analysis was performed to compare the image processed equivalent diameter (ED), inter-fibrillar distance (IFD), and EFD of the CFs of treated versus normal corneas. The ReNu-soaked cornea resulted in partly degenerated epithelium with loose hemidesmosomes and Bowman's collagen. In contrast, the epithelium of the cornea soaked in Hippia was degenerated or lost but showed closely packed Bowman's collagen. Soaking the corneas in both MPS caused a statistically significant decrease in the anterior collagen fibril, ED and a significant change in IFD, and EFD than those of the untreated corneas (p<0.05, for all comparisons). The introduction of EFD measurement in the study directly provided a sense of gap between periphery of the collagen bundles, their spatial distribution; and in combination with ED, they showed how the corneal collagen bundles are spaced in relation to their diameters. The spatial distribution parameter NSDEFD indicated that ReNu treated cornea fibrils were uniformly distributed spatially, followed by normal and Hippia. The EFD measurement with relatively lower standard deviation and NSDEFD, a characteristic of uniform CFs distribution, can be an additional parameter used in evaluating collagen organization and accessing the effects of various treatments on corneal health and transparency. Copyright © 2015 Elsevier Ltd. All rights reserved.

Multiparameter thermo-mechanical OCT-based characterization of laser-induced cornea reshaping

NASA Astrophysics Data System (ADS)

Zaitsev, Vladimir Yu.; Matveyev, Alexandr L.; Matveev, Lev A.; Gelikonov, Grigory V.; Vitkin, Alex; Omelchenko, Alexander I.; Baum, Olga I.; Shabanov, Dmitry V.; Sovetsky, Alexander A.; Sobol, Emil N.

2017-02-01

Phase-sensitive optical coherence tomography (OCT) is used for visualizing dynamic and cumulative strains and corneashape changes during laser-produced tissue heating. Such non-destructive (non-ablative) cornea reshaping can be used as a basis of emerging technologies of laser vision correction. In experiments with cartilaginous samples, polyacrilamide phantoms and excised rabbit eyes we demonstrate ability of the developed OCT system to simultaneously characterize transient and cumulated strain distributions, surface displacements, scattering tissue properties and possibility of temperature estimation via thermal-expansion measurements. The proposed approach can be implemented in perspective real-time OCT systems for ensuring safety of new methods of laser reshaping of cornea.

Quantization of collagen organization in the stroma with a new order coefficient

PubMed Central

Germann, James A.; Martinez-Enriquez, Eduardo; Marcos, Susana

2017-01-01

Many optical and biomechanical properties of the cornea, specifically the transparency of the stroma and its stiffness, can be traced to the degree of order and direction of the constituent collagen fibers. To measure the degree of order inside the cornea, a new metric, the order coefficient, was introduced to quantify the organization of the collagen fibers from images of the stroma produced with a custom-developed second harmonic generation microscope. The order coefficient method gave a quantitative assessment of the differences in stromal collagen arrangement across the cornea depths and between untreated stroma and cross-linked stroma. PMID:29359095

21 CFR 524.1982 - Proparacaine ophthalmic solution.

Code of Federal Regulations, 2014 CFR

2014-04-01

... is used as an anesthetic in cauterization of corneal ulcers, removal of foreign bodies and sutures from the cornea, and measurement of intraocular pressure (tonometry) when glaucoma is suspected; as an... cauterization of corneal ulcers, removal of foreign bodies and sutures from the cornea, and measurement of...

Establishment of a new in vitro test method for evaluation of eye irritancy using a reconstructed human corneal epithelial model, LabCyte CORNEA-MODEL.

PubMed

Katoh, Masakazu; Hamajima, Fumiyasu; Ogasawara, Takahiro; Hata, Ken-ichiro

2013-12-01

Finding in vitro eye irritation testing alternatives to animal testing such as the Draize eye test, which uses rabbits, is essential from the standpoint of animal welfare. It has been developed a reconstructed human corneal epithelial model, the LabCyte CORNEA-MODEL, which has a representative corneal epithelium-like structure. Protocol optimization (pre-validation study) was examined in order to establish a new alternative method for eye irritancy evaluation with this model. From the results of the optimization experiments, the application periods for chemicals were set at 1min for liquid chemicals or 24h for solid chemicals, and the post-exposure incubation periods were set at 24h for liquids or zero for solids. If the viability was less than 50%, the chemical was judged to be an eye irritant. Sixty-one chemicals were applied in the optimized protocol using the LabCyte CORNEA-MODEL and these results were evaluated in correlation with in vivo results. The predictions of the optimized LabCyte CORNEA-MODEL eye irritation test methods were highly correlated with in vivo eye irritation (sensitivity 100%, specificity 80.0%, and accuracy 91.8%). These results suggest that the LabCyte CORNEA-MODEL eye irritation test could be useful as an alternative method to the Draize eye test. Copyright © 2013 Elsevier Ltd. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Akhtar, R.A.; Abdel-Latif, A.A.

Muscarinic cholinergic and ..cap alpha../sub 1/-adrenergic agonists provoke hydrolysis of PIP/sub 2/ into diacylglycerol (DG) and inositol trisphosphate (IP/sub 3/) in a wide variety of tissue. Recently, IP/sub 3/ has been shown to mobilize Ca/sup 2 +/ from ER in several permeabilized tissue preparations. Although rabbit cornea is enriched in ACh and NE, the physiological function of these neurotransmitters is unclear. The present studies were initiated to determine the effects of cholinergic and adrenergic agonists on PIP/sub 2/ turnover in the cornea. Addition of ACh or NE (50 ..mu..M each) to the /sup 32/P-labeled corneas for 10 min decreased themore » radioactivity in PIP/sub 2/ by 33 and 36%, and increased the radioactivity in phosphatidic acid by 72 and 52%, respectively. When the corneas were labeled with myo-(/sup 3/H)inositol, ACh and NE increased the accumulation of IP/sub 3/ by 92 and 48%, respectively. The effects of ACh and NE on phospholipid labeling and IP/sub 3/ accumulation were specifically inhibited by atropine (10 ..mu..M) and prazosin (10 ..mu..M), respectively. The data suggest the presence of muscarinic cholinergic and ..cap alpha../sub 1/-adrenergic receptors in the rabbit cornea. Furthermore, activation of these receptors leads to cleavage of PIP/sub 2/ into DG and IP/sub 3/ which may function as second messengers in this tissue.« less

Numerical model and analysis of an energy-based system using microwaves for vision correction

NASA Astrophysics Data System (ADS)

Pertaub, Radha; Ryan, Thomas P.

2009-02-01

A treatment system was developed utilizing a microwave-based procedure capable of treating myopia and offering a less invasive alternative to laser vision correction without cutting the eye. Microwave thermal treatment elevates the temperature of the paracentral stroma of the cornea to create a predictable refractive change while preserving the epithelium and deeper structures of the eye. A pattern of shrinkage outside of the optical zone may be sufficient to flatten the central cornea. A numerical model was set up to investigate both the electromagnetic field and the resultant transient temperature distribution. A finite element model of the eye was created and the axisymmetric distribution of temperature calculated to characterize the combination of controlled power deposition combined with surface cooling to spare the epithelium, yet shrink the cornea, in a circularly symmetric fashion. The model variables included microwave power levels and pulse width, cooling timing, dielectric material and thickness, and electrode configuration and gap. Results showed that power is totally contained within the cornea and no significant temperature rise was found outside the anterior cornea, due to the near-field design of the applicator and limited thermal conduction with the short on-time. Target isothermal regions were plotted as a result of common energy parameters along with a variety of electrode shapes and sizes, which were compared. Dose plots showed the relationship between energy and target isothermic regions.

Effects of ablation depth and repair time on the corneal elastic modulus after laser in situ keratomileusis.

PubMed

Wang, Xiaojun; Li, Xiaona; Chen, Weiyi; He, Rui; Gao, Zhipeng; Feng, Pengfei

2017-01-17

The biomechanical properties of the cornea should be taken into account in the refractive procedure in order to perform refractive surgery more accurately. The effects of the ablation depth and repair time on the elastic modulus of the rabbit cornea after laser in situ keratomileusis (LASIK) are still unclear. In this study, LASIK was performed on New Zealand rabbits with different ablation depth (only typical LASIK flaps were created; residual stroma bed was 50 or 30% of the whole cornea thickness respectively). The animals without any treatment were served as normal controls. The corneal thickness was measured by ultrasonic pachymetry before animals were humanly killed after 7 or 28 days post-operatively. The corneal elastic modulus was measured by uniaxial tensile testing. A mathematical procedure considering the actual geometrics of the cornea was created to analyze the corneal elastic modulus. There were no obvious differences among all groups in the elastic modulus on after 7 days post-operatively. However, after 28th days post-operatively, there was a significant increase in the elastic modulus with 50 and 30% residual stroma bed; only the elastic modulus of the cornea with 30% residual stroma bed was significantly higher than that of 7 days. Changes in elastic modulus after LASIK suggest that this biomechanical effect may correlate with the ablation depth and repair time.

In vivo imaging through the entire thickness of human cornea by full-field optical coherence tomography

NASA Astrophysics Data System (ADS)

Mazlin, Viacheslav; Xiao, Peng; Dalimier, Eugénie; Grieve, Kate; Irsch, Kristina; Sahel, José; Fink, Mathias; Boccara, Claude

2018-02-01

Despite obvious improvements in visualization of the in vivo cornea through the faster imaging speeds and higher axial resolutions, cellular imaging stays unresolvable task for OCT, as en face viewing with a high lateral resolution is required. The latter is possible with FFOCT, a method that relies on a camera, moderate numerical aperture (NA) objectives and an incoherent light source to provide en face images with a micrometer-level resolution. Recently, we for the first time demonstrated the ability of FFOCT to capture images from the in vivo human cornea1. In the current paper we present an extensive study of appearance of healthy in vivo human corneas under FFOCT examination. En face corneal images with a micrometer-level resolution were obtained from the three healthy subjects. For each subject it was possible to acquire images through the entire corneal depth and visualize the epithelium structures, Bowman's layer, sub-basal nerve plexus (SNP) fibers, anterior, middle and posterior stroma, endothelial cells with nuclei. Dimensions and densities of the structures visible with FFOCT, are in agreement with those seen by other cornea imaging methods. Cellular-level details in the images obtained together with the relatively large field-of-view (FOV) and contactless way of imaging make this device a promising candidate for becoming a new tool in ophthalmological diagnostics.

Automated lamellar therapeutic keratoplasty with fibrin adhesive in the treatment of anterior corneal opacities.

PubMed

Hashemi, Hassan; Dadgostar, Alhan

2011-06-01

To assess the visual outcome of using fibrin adhesive in automated lamellar therapeutic keratoplasty with a microkeratome in the treatment of anterior corneal opacities. In this prospective noncomparative clinical trial, surgery was done on 10 eyes belonging to 9 patients with anterior stromal opacity (macular dystrophy, spheroidal degeneration, scarring because of advanced recurrent pterygium, refractive surgery, or trauma). Depending on the depth of the opacity, a 130- or 250-μm flap was removed from the recipient cornea using a microkeratome. Then, a thin layer of fibrin adhesive was spread over the bed, and a lenticule with the same thickness, created from the donor cornea, was positioned in place. After allowing the glue to set for about 5 minutes, a bandage contact lens was placed over the cornea, which was removed 7-10 days postoperatively. All corneas healed properly, and none required suturing or reoperation. During the follow-up period, no inflammation or rejection was observed. The donor cornea and the donor-recipient interface remained clear in all cases. The mean of best contact lens-corrected visual acuity improved from 1.14 ± 0.53 to 0.51 ± 0.23 in the logarithm of the minimum angle of resolution scale. The fibrin glue can provide safe and effective attachment needed in automated lamellar therapeutic keratectomy and obviates the need for suturing. However, it requires improvement for easier and safer use in ophthalmology.

Thick keratoconic cornea associated with posterior polymorphous corneal dystrophy.

PubMed

Zaarour, K; Slim, E; Antoun, J; Waked, N

2017-03-01

We herein report a case of bilateral unusually thick non-edematous keratoconic corneas with associated endothelial features of posterior polymorphous corneal dystrophy (PPCD). We report the case of a 27-year-old myopic woman who presented for refractive surgery. Slit lamp exam showed bilateral corneal protrusion with diffuse deep stromal and endothelial vesicular opacities and small paracentral bands. Topography showed generalized advanced corneal steepening in both eyes with increased anterior and posterior central corneal elevations in comparison to the best fit sphere. Ultrasound pachymetry showed central corneal thickness of 605μm (RE) and 612μm (LE). On specular biomicroscopy, cell density of 2503 cells/mm 2 RE and 1526 cells/mm 2 LE with significant cellular pleomorphism and polymegathism were noted. Clinical and paraclinical findings together suggest the presence of simultaneous keratoconus and PPCD. The literature has suggested an association between PPCD and steep cornea. Moreover, many reports have also described cases of associated PPCD and keratoconus with characteristic thinning and ectasia, in comparison to the unusual thick corneas noted in our patient, despite the absence of edema. Identification of genetics factors is further needed to clarify this association. This case describes a patient whose corneas present features of both keratoconus and PPCD and is unique due to the presence of increased corneal thickness despite the absence of edema. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

The tissue-engineered human cornea as a model to study expression of matrix metalloproteinases during corneal wound healing.

PubMed

Couture, Camille; Zaniolo, Karine; Carrier, Patrick; Lake, Jennifer; Patenaude, Julien; Germain, Lucie; Guérin, Sylvain L

2016-02-01

Corneal injuries remain a major cause of consultation in the ophthalmology clinics worldwide. Repair of corneal wounds is a complex mechanism that involves cell death, migration, proliferation, differentiation, and extracellular matrix (ECM) remodeling. In the present study, we used a tissue-engineered, two-layers (epithelium and stroma) human cornea as a biomaterial to study both the cellular and molecular mechanisms of wound healing. Gene profiling on microarrays revealed important alterations in the pattern of genes expressed by tissue-engineered corneas in response to wound healing. Expression of many MMPs-encoding genes was shown by microarray and qPCR analyses to increase in the migrating epithelium of wounded corneas. Many of these enzymes were converted into their enzymatically active form as wound closure proceeded. In addition, expression of MMPs by human corneal epithelial cells (HCECs) was affected both by the stromal fibroblasts and the collagen-enriched ECM they produce. Most of all, results from mass spectrometry analyses provided evidence that a fully stratified epithelium is required for proper synthesis and organization of the ECM on which the epithelial cells adhere. In conclusion, and because of the many characteristics it shares with the native cornea, this human two layers corneal substitute may prove particularly useful to decipher the mechanistic details of corneal wound healing. Copyright © 2015 Elsevier Ltd. All rights reserved.

Comparison of genome-wide gene expression in suture- and alkali burn-induced murine corneal neovascularization

PubMed Central

Jia, Changkai; Zhu, Wei; Ren, Shengwei; Xi, Haijie; Li, Siyuan

2011-01-01

Purpose Suture placement and alkali burn to the cornea are often used to induce inflammatory corneal neovascularization (CorNV) models in animals. This study compares the changes in genome-wide gene expression under these two CorNV conditions in mice. Methods CorNV were induced in Balb/c mice by three interrupted 10–0 sutures placed at sites about 1 mm from the corneal apex, or by alkali burns that were 2 mm in size in the central area of the cornea. At the points in time when neovascularization progressed most quickly, some eyeballs were subjected to histological staining to examine CorNV and inflammatory cells infiltration, and some corneas were harvested to extract mRNA for microarray assay. After normalization and filtering, the microarray data were subject to statistical analysis using Significance Analysis of Microarray software, and interested genes were annotated using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) program. The expression change of classical proangiogenic molecule like vascular endothelial growth factor (VEGF) and antiangiogenic molecule like pigment epithelium-derived factor (PEDF) was further verified using western blotting. Results Suture placement induced CorNV in the areas between the suture and limbus, but did not affect the transparency of the yet unvasuclarized areas of the corneas. In contrast, alkali burn caused edema and total loss of transparency of the whole cornea. Histology showed that sutures only caused localized epithelial loss and inflammatory infiltration between the suture and limbus, but chemical burn depleted the whole epithelial layer of the central cornea and caused heavy cellular infiltration of the whole cornea. At day 5 after suture placement, 1,055 differentially expressed probes were identified, out of which 586 probes were upregulated and 469 probes were downregulated. At a comparable time point, namely on day 6 after the alkali burn to the corneas, 472 probes were upregulated and 389 probes were downregulated. Among these differentially expressed probes, a significant portion (530 probes in total, including 286 upregulated and 244 downregulated probes) showed a similar pattern of change in both models. Annotation (using DAVID) of the overlapping differential genes revealed that the significant enrichment gene ontology terms were “chemotaxis” and “immune response” for the upregulated genes, and “oxidation reduction” and “programmed cell death” for the downregulated genes. Some genes or gene families (e.g., S100A family or α-, β-, or γ-crystallin family) that had not been related to corneal pathogenesis or neovascularization were also revealed to be involved in CorNV. VEGF was upregulated and PEDF was stable as shown with western blotting. Conclusions Sutures and alkali burn to the corneas produced types of damage that affected transparency differentially, but gene profiling revealed similar patterns of changes in gene expression in these two CorNV models. Further studies of the primary genes found to be involved in CorNV will supplement current understanding about the pathogenesis of neovascularization diseases. PMID:21921991

A simplified technique for in situ excision of cornea and evisceration of retinal tissue from human ocular globe.

PubMed

Parekh, Mohit; Ferrari, Stefano; Di Iorio, Enzo; Barbaro, Vanessa; Camposampiero, Davide; Karali, Marianthi; Ponzin, Diego; Salvalaio, Gianni

2012-06-12

Enucleation is the process of retrieving the ocular globe from a cadaveric donor leaving the rest of the globe undisturbed. Excision refers to the retrieval of ocular tissues, especially cornea, by cutting it separate from the ocular globe. Evisceration is the process of removing the internal organs referred here as retina. The ocular globe consists of the cornea, the sclera, the vitreous body, the lens, the iris, the retina, the choroid, muscles etc (Suppl. Figure 1). When a patient is suffering from corneal damage, the cornea needs to be removed and a healthy one must be transplanted by keratoplastic surgeries. Genetic disorders or defects in retinal function can compromise vision. Human ocular globes can be used for various surgical procedures such as eye banking, transplantation of human cornea or sclera and research on ocular tissues. However, there is little information available on human corneal and retinal excision, probably due to the limited accessibility to human tissues. Most of the studies describing similar procedures are performed on animal models. Research scientists rely on the availability of properly dissected and well-conserved ocular tissues in order to extend the knowledge on human eye development, homeostasis and function. As we receive high amount of ocular globes out of which approximately 40% (Table 1) of them are used for research purposes, we are able to perform huge amount of experiments on these tissues, defining techniques to excise and preserve them regularly. The cornea is an avascular tissue which enables the transmission of light onto the retina and for this purpose should always maintain a good degree of transparency. Within the cornea, the limbus region, which is a reservoir of the stem cells, helps the reconstruction of epithelial cells and restricts the overgrowth of the conjunctiva maintaining corneal transparency and clarity. The size and thickness of the cornea are critical for clear vision, as changes in either of them could lead to distracted, unclear vision. The cornea comprises of 5 layers; a) epithelium, b) Bowman's layer, c) stroma, d) Descemet's membrane and e) endothelium. All layers should function properly to ensure clear vision(4,5,6). The choroid is the intermediate tunic between the sclera and retina, bounded on the interior by the Bruch's membrane and is responsible for blood flow in the eye. The choroid also helps to regulate the temperature and supplies nourishment to the outer layers of the retina(5,6). The retina is a layer of nervous tissue that covers the back of the ocular globe (Suppl. Figure 1) and consists of two parts: a photoreceptive part and a non-receptive part. The retina helps to receive the light from the cornea and lens and converts it into the chemical energy eventually transmitted to the brain with help of the optic nerve(5,6). The aim of this paper is to provide a protocol for the dissection of corneal and retinal tissues from human ocular globes. Avoiding cross-contamination with adjacent tissues and preserving RNA integrity is of fundamental importance as such tissues are indispensable for research purposes aimed at (i) characterizing the transcriptome of the ocular tissues, (ii) isolating stem cells for regenerative medicine projects, and (iii) evaluating histological differences between tissues from normal/affected subjects. In this paper we describe the technique we currently use to remove the cornea, the choroid and retinal tissues from an ocular globe. Here we provide a detailed protocol for the dissection of the human ocular globe and the excision of corneal and retinal tissues. The accompanying video will help researchers to learn an appropriate technique for the retrieval of precious human tissues which are difficult to find regularly.

Removal of a barbed fish hook from the cornea of an 8-year-old boy.

PubMed

Kemp, Pavlina S; Shah, Ankoor S; Elliott, Alexandra T; Wan, Michael J

2017-10-01

This video article shows the surgical removal of a three-pronged barbed fish hook from the cornea and anterior chamber of an 8-year-old boy. The advance-and-cut method is shown: the embedded barbed point of the fish hook was advanced through the cornea, its point was cut off, and the shank of the hook was withdrawn. This method was chosen due to the three-pronged configuration of the fish hook to minimize additional trauma to the eye. Other methods of fish hook removal are briefly reviewed. Copyright © 2017 American Association for Pediatric Ophthalmology and Strabismus. Published by Elsevier Inc. All rights reserved.

Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope

NASA Astrophysics Data System (ADS)

Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

2016-03-01

Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope.

PubMed

Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

2016-03-01

Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

Three-Dimensional Ultrasonic Imaging Of The Cornea

NASA Technical Reports Server (NTRS)

Heyser, Rrichar C.; Rooney, James A.

1988-01-01

Proposed technique generates pictures of curved surfaces. Object ultrasonically scanned in raster pattern generated by scanning transmitter/receiver. Receiver turned on at frequent intervals to measure depth variations of scanned object. Used for medical diagnoses by giving images of small curved objects as cornea. Adaptable to other types of reflection measurementsystems such as sonar and radar.

[Corneal transparency: anatomical basis and evaluation methods].

PubMed

Avetisov, S E; Narbut, M N

Being just a relatively small part of the fibrous tunic of the eyeball, the cornea is, nevertheless, unique in terms of the variety of its functions. Because the cornea differs significantly from other protective frames in its structure, it provides the possibility of light transmission and strong refraction that largely contributes to the total refraction of the eye. The development of ophthalmology is impossible without improving methods of adequate anatomical and functional assessment of the eye not only as a whole, but also as a collection of interacting structures.In this regard, examination methods of the cornea have undergone significant advances in recent years. So far, the level of corneal transparency has been judged by biomicroscopy findings or indirect characteristics (thickness, structure, etc.). Confocal microscopy of the cornea and wave-based examinations involving one of the available laser interferometers (OCT or HRT) are also used. However, the data obtained with these methods resembles that of layer-specific reflectometry, i.e. the magnitude of directed reflection of the light beam from corneal corpuscles, which does not completely agree with the classical idea of transparency.

In-vivo measurements of the tear film on a cornea and a contact lens by use of interferometry

NASA Astrophysics Data System (ADS)

Licznerski, Tomasz J.; Kasprzak, Henryk T.; Kowalik, Waldemar

1996-12-01

The tear film fulfills several important functions in the eye. Apart of its physiologic functions like maintaining a moist environment for the epithelial cells of the cornea and conjunctiva, bacterial properties, transporting metabolic products etc., this film causes that the corneal surface has the optical quality. This smooth surface allows to apply interferometry for measurements. The paper presents tear's layer distribution on the soft contact lens and the cornea in comparison. Tv frame speed registration in the Twyman- Green interferometer was used to observe an unstable biomedical objects like the eye. The proposed method has the advantage of being noncontact and applies the low energy laser beam in interferometric set-up. This provides non- invasive testing of human cornea in vivo and enables observation the kinetics of its tear layer deterioration. The evaluation of non-invasive tear breakup time is possible by use of proposed setup. Further analysis of recorded interferograms helps to examine the matter of the breakup process and can be used for detection of the 'dry eye' symptoms.

Diffusion and Monod kinetics model to determine in vivo human corneal oxygen-consumption rate during soft contact lens wear

PubMed Central

Del Castillo, Luis F.; da Silva, Ana R. Ferreira; Hernández, Saul I.; Aguilella, M.; Andrio, Andreu; Mollá, Sergio; Compañ, Vicente

2014-01-01

Purpose We present an analysis of the corneal oxygen consumption Qc from non-linear models, using data of oxygen partial pressure or tension (pO2) obtained from in vivo estimation previously reported by other authors.1 Methods Assuming that the cornea is a single homogeneous layer, the oxygen permeability through the cornea will be the same regardless of the type of lens that is available on it. The obtention of the real value of the maximum oxygen consumption rate Qc,max is very important because this parameter is directly related with the gradient pressure profile into the cornea and moreover, the real corneal oxygen consumption is influenced by both anterior and posterior oxygen fluxes. Results Our calculations give different values for the maximum oxygen consumption rate Qc,max, when different oxygen pressure values (high and low pO2) are considered at the interface cornea-tears film. Conclusion Present results are relevant for the calculation on the partial pressure of oxygen, available at different depths into the corneal tissue behind contact lenses of different oxygen transmissibility. PMID:25649636

SVM-based automatic diagnosis method for keratoconus

NASA Astrophysics Data System (ADS)

Gao, Yuhong; Wu, Qiang; Li, Jing; Sun, Jiande; Wan, Wenbo

2017-06-01

Keratoconus is a progressive cornea disease that can lead to serious myopia and astigmatism, or even to corneal transplantation, if it becomes worse. The early detection of keratoconus is extremely important to know and control its condition. In this paper, we propose an automatic diagnosis algorithm for keratoconus to discriminate the normal eyes and keratoconus ones. We select the parameters obtained by Oculyzer as the feature of cornea, which characterize the cornea both directly and indirectly. In our experiment, 289 normal cases and 128 keratoconus cases are divided into training and test sets respectively. Far better than other kernels, the linear kernel of SVM has sensitivity of 94.94% and specificity of 97.87% with all the parameters training in the model. In single parameter experiment of linear kernel, elevation with 92.03% sensitivity and 98.61% specificity and thickness with 97.28% sensitivity and 97.82% specificity showed their good classification abilities. Combining elevation and thickness of the cornea, the proposed method can reach 97.43% sensitivity and 99.19% specificity. The experiments demonstrate that the proposed automatic diagnosis method is feasible and reliable.

Characterization of elasticity and hydration of composite hydrogel based on collagen-iota carrageenan as a corneal tissue engineering

NASA Astrophysics Data System (ADS)

Rinawati, M.; Triastuti, J.; Pursetyo, K. T.

2018-04-01

The cornea is a refractive element of the eye that serves to continue the stimulation of light into the eye it has a clear, transparent, elastic and relatively thick tissue. Factors caused corneal blindness, are dystrophy, keratoconus, corneal scaring. Hydrogels can be made from polysaccharide derivatives that have gelation properties such as iota carrageenan. Therefore, it is a need to develop composite hydrogel based collagen-iota carragenan as an engineeried corneal tissue with high elasticity and hydration properties. Collagen hydrogel has a maximum water content an has equlibrium up to 40 %, less than the human cornea, 81 % and under normal hydration conditions, the human cornea can transmit 87 % of visible light. In addition, the refractive index on the surface of the cornea with air is 1.375-1.380. Based on this study, it is necessary to conduct research on the development and composition of hydrogel composite collagen-iota carrageen hydrogen based on. The best result was K5 (5:5) treatment, which has the equilibrium water content of 87.07 % and viscosity of 10.7346 Pa.s.

Importance of accurately assessing biomechanics of the cornea.

PubMed

Roberts, Cynthia J

2016-07-01

This article summarizes the state-of-the-art in clinical corneal biomechanics, including procedures in which biomechanics play a role, and the clinical consequences in terms of error in estimating intraocular pressure (IOP). Corneal biomechanical response to refractive surgery can be categorized into either stable alteration of surface shape and thus visual outcome, or unstable biomechanical decompensation. The stable response is characterized by central flattening and peripheral steepening that is potentiated in a stiffer cornea. Two clinical devices for assessing corneal biomechanics do not yet measure classic biomechanical properties, but rather provide assessment of corneal deformation response. Biomechanical parameters are a function of IOP, and both the cornea and sclera become stiffer as IOP increases. Any assessment of biomechanical parameters must include IOP, and one value of stiffness does not adequately characterize a cornea. Corneal biomechanics plays a role in the outcomes of any procedure in which lamellae are transected. Once the corneal structure has been altered in a manner that includes central thinning, IOP measurements with applanation tonometry are likely not valid, and other technologies should be used.

The influence of viewing an entertainment-education program on cornea donation intention: a test of the theory of planned behavior.

PubMed

Bae, Hyuhn-Suhck; Kang, Seok

2008-01-01

This study was designed to examine the role of issue involvement and the 3 components of the theory of planned behavior in predicting intentions to sign a cornea donor card. This study also compares viewers and nonviewers of an entertainment-education program in terms of issue involvement and the theory of planned behavior. The breakdown of effects for the latent constructs confirms that issue involvement is an important intermediary in the persuasion process. Issue involvement is a common causal antecedent of attitude, subjective norm, and perceived behavioral control, the last 2 of which, in turn, predict intentions, whereas attitude does not. The revised path model confirms that involvement directly influences intentions. In the comparison of viewers with nonviewers, viewers exhibit a significantly higher degree of involvement, attitude toward cornea donation, subjective norm, perceived behavioral control, and intention to sign a cornea donor card. The overall findings of this study suggest that adding issue involvement in the theory of planned behavior enhances the explanatory power of the theory in predicting intentions.

Role of innate and adaptive immunity in the pathogenesis of keratitis.

PubMed

Hazlett, Linda D

2005-01-01

Pseudomonas aeruginosa is a common organism associated with bacterial keratitis primarily resulting from contact lens usage. Advances in our understanding of host innate and adaptive immune responses to experimental infection have been achieved using animal models, including inbred mouse models that are classed as resistant (cornea heals) vs. susceptible (cornea perforates). Evidence has shown that sustained IL-12-driven IFN-gamma production in dominant Th1 responder strains such as C57BL/6 (B6) contributes to corneal destruction and perforation. In contrast, in Th2-responder BALB/c mice, IL-18-driven IFN-gamma production regulates bacterial killing with less corneal destruction. IL-1 and chemotactic cytokines (e.g., MIP-2) recruit PMN to the cornea. The critical role of these cells in the innate immune response and their regulation after bacterial infection has been established. The studies provide a better understanding of the regulatory mechanisms that operate in the cornea after P. aeruginosa challenge, determining susceptibility vs. resistance to disease, and are consistent with long-term goals of providing targets for better treatment of disease.

Real-time analysis keratometer

NASA Technical Reports Server (NTRS)

Adachi, Iwao P. (Inventor); Adachi, Yoshifumi (Inventor); Frazer, Robert E. (Inventor)

1987-01-01

A computer assisted keratometer in which a fiducial line pattern reticle illuminated by CW or pulsed laser light is projected on a corneal surface through lenses, a prismoidal beamsplitter quarterwave plate, and objective optics. The reticle surface is curved as a conjugate of an ideal corneal curvature. The fiducial image reflected from the cornea undergoes a polarization shift through the quarterwave plate and beamsplitter whereby the projected and reflected beams are separated and directed orthogonally. The reflected beam fiducial pattern forms a moire pattern with a replica of the first recticle. This moire pattern contains transverse aberration due to differences in curvature between the cornea and the ideal corneal curvature. The moire pattern is analyzed in real time by computer which displays either the CW moire pattern or a pulsed mode analysis of the transverse aberration of the cornea under observation, in real time. With the eye focused on a plurality of fixation points in succession, a survey of the entire corneal topography is made and a contour map or three dimensional plot of the cornea can be made as a computer readout in addition to corneal radius and refractive power analysis.

Drug-induced corneal damage.

PubMed

2014-04-01

Corneal damage can have a variety of causes, including infections, chemical splashes, environmental factors (radiation, trauma, contact lenses, etc.), and systemic diseases (genetic, autoimmune, inflammatory, metabolic, etc.). A wide range of drugs can also damage the cornea. The severity of drug-induced corneal changes can range from simple asymptomatic deposits to irreversible, sight-threatening damage. Several factors can influence the onset of corneal lesions. Some factors, such as the dose, are treatment-related, while others such as contact lenses, are patient-related. A variety of mechanisms may be involved, including corneal dryness, changes in the corneal epithelium, impaired wound healing and deposits. Many drugs can damage the cornea through direct contact, after intraocular injection or instillation, including VEGF inhibitors, anti-inflammatory drugs, local anaesthetics, glaucoma drugs, fluoroquinolones, and preservatives. Some systemically administered drugs can also damage the cornea, notably cancer drugs, amiodarone and isotretinoin. Vulnerable patients should be informed of this risk if they are prescribed a drug with the potential to damage the cornea so that they can identify problems in a timely manner. It may be necessary to discontinue the suspect drug when signs and symptoms of corneal damage occur.

The Resin-Embedded Cornea Prepared Via Rapid Processing Protocol : A Good Histomorphometric Target for Clinical Investigation in Ophthalmology and Optometry

PubMed Central

Cheah, Pike See; Mohidin, Norhani; Mohd Ali, Bariah; Maung, Myint; Latif, Azian Abdul

2008-01-01

This study illustrates and quantifies the changes on corneal tissue between the paraffin-embedded and resin-embedded blocks and thus, selects a better target in investigational ophthalmology and optometry via light microscopy. Corneas of two cynomolgus monkeys (Macaca fascicularis) were used in this study. The formalin-fixed cornea was prepared in paraffin block via the conventional tissue processing protocol (4-day protocol) and stained with haematoxylin and eosin. The glutaraldehyde-fixed cornea was prepared in resin block via the rapid and modified tissue processing procedure (1.2-day protocol) and stained with toluidine blue. The paraffin-embedded sample exhibits various undesired tissue damage and artifact such as thinner epithelium (due to the substantial volumic extraction from the tissue), thicker stroma layer (due to the separation of lamellae and the presence of voids) and the distorted endothelium. In contrast, the resin-embedded corneal tissue has demonstrated satisfactory corneal ultrastructural preservation. The rapid and modified tissue processing method for preparing the resin-embedded is particularly beneficial to accelerate the microscopic evaluation in ophthalmology and optometry. PMID:22570589

Ultrasonic biomicroscopy in ophthalmology and eye banking

NASA Astrophysics Data System (ADS)

Rosenwasser, George O. D.

1999-06-01

Echography has become a valuable diagnostic tool in ophthalmology. Ultrasonic biomicroscopy (UBM) in particular may be applied to the evaluation of small lesions of the anterior segment of the eye. Disease processes such as conjunctival and iris melanoma, other forms of neoplasia, intraocular cysts, narrow angle glaucoma, and intraocular foreign bodies can be diagnostically evaluated and followed longitudinally by UBM. Combining UBM with spectroscopy may become useful in determining cell type origins of a variety of tumors. Eye banking also has an increased need for UBM in corneal tissue banking. The recent development of the Laser In Situ Keratomileusis procedure has allowed corneal surgeries to create a partial thickness flap of tissue in the cornea, remove tissue from the base of the cornea with excimer laser ablation, and replace the hinged flap. This causes a substantial change in refractive error while thinning the cornea and leaving an interface within the corneal stroma. The ability to detect this type of surgery is essential in eye banking. Ultrasonic pachymetry to determine central thickness and biomicroscopy to detect the presence of an interface are essential in avoiding the use of these corneas for transplantation purposes. Determining the topography of the preserved corneas is another potential application for ultrasonography. Using this information to reduce optical aberration after transplant is crucial in improving visual performance post transplantation. A review of the anatomy of the eye, pathology of ocular diseases relevant to UBM, and principles of eye banking will be presented.

In-vitro corneal transparency measuring system

NASA Astrophysics Data System (ADS)

Ventura, Liliane; da Costa Vieira, Marcelo A.; Isaac, Flavio; Chiaradia, Caio; Faria de Sousa, Sidney J.

2001-06-01

A system for measuring the average corneal transparency of preserved corneas has been developed in order to provide a more accurate and standard report of the corneal tissue. The donated cornea transparency is one of the features to be analyzed previously to its indication for the transplant. The small portable system consists of two main parts: the optical and the electronic parts. The optical system consists of a white light, lenses and pin-holes that collimate white light beams that illuminates the cornea in its preservative medium. The light that passes through the cornea is detected by a resistive detector and the average corneal transparency is shown in a display. In order to obtain just the tissue transparency, the electronic circuit was built in a way that there is a baseline input of the preservative medium, previous to the measurement of the corneal transparency. Manipulating the system consists of three steps: (1) Adjusting the zero percentage in the absence of light (at this time the detectors in the dark); (2) Placing the preservative medium in the system and adjusting the 100% value (this is the baseline input); (3) Preserving the cornea and placing it in the system. The system provides the tissue transparency. The system is connected to an endothelium evaluation system for Slit Lamp, that we have developed, and statistics about the relationship of the corneal transparency and density of the endothelial cells will be provided in the next years. The system is being used in a public Eye Bank in Brasil.

Study of light scattering and transparency in human edematous corneas and application to corneal grafts

NASA Astrophysics Data System (ADS)

Marciano, Tal; Peyrot, Donald; Crotti, Caroline; Alahyane, Fatima; Kowalczuk, Laura; Plamann, Karsten

2011-07-01

The optical properties of the cornea have been a research subject of great interest for many years. Several early theories have been put forward to explain with more or less success the optical transparency of this tissue, but it was not until Maurice demonstrated in a very elegant way during the 50s that this optical transparency could be explained by the regular ultrastructure of the cornea. When becoming edematous, the cornea's ultrastructure is perturbed and the tissue becomes a strongly scattering medium. With the emergence of ophthalmologic surgery by ultrashort pulse lasers in recent years, a regain of interest in the subject of corneal transparency arose. However, relatively little and no recent data of transparency spectra measurements covering a large wavelength range is available in the literature. The purpose of this study is to provide quantitative values for light scattering and its relation to the degree of edema by measuring the spectrum of transmitted light through corneas presenting different degrees of edema. This paper focus on the comparison of laboratory measurements published earlier with a new simple method we propose We also for eye banks to quantitatively measure the degree of transparency of corneal grafts by measuring the modulation transfer function of a Siemens star viewed through a corneal graft. Indeed, there is no current method to determine the transparency of corneal graft but the subjectivity of the laboratory technician or the ophthalmic surgeon.

Aspergillus fumigatus Increased PAR-2 Expression and Elevated Proinflammatory Cytokines Expression Through the Pathway of PAR-2/ERK1/2 in Cornea.

PubMed

Niu, Yawen; Zhao, Guiqiu; Li, Cui; Lin, Jing; Jiang, Nan; Che, Chengye; Zhang, Jie; Xu, Qiang

2018-01-01

To determine the role of protease-activated receptor-2 (PAR-2) in cornea infected by Aspergillus fumigatus. PAR-2 was tested in normal and infected corneas of C57BL/6 mice. Mice corneas were infected with A. fumigatus with or without pretreatment of PAR-2 antagonist (FSLLRY-NH2). Polymorphonuclear neutrophilic leukocytes (PMNs) were stimulated with 75% ethanol-killed A. fumigatus with or without pretreatment of FSLLRY-NH2. Disease severity was documented by clinical score and photographs with a slit lamp. PCR, Western blot, and ELISA tested expression of PAR-2, IL-1β, TNF-α, IFN-γ, MIP-2, and p-ERK1/2. PMN infiltration was assessed by myeloperoxidase assay and immunofluorescent staining. PAR-2 expression was significantly elevated by A. fumigatus, whereas the upregulation was significantly inhibited by FSLLRY-NH2 in mice corneas. FSLLRY-NH2 decreased disease response, PMN infiltration, and proinflammatory cytokine expression compared with infected control. In PMNs, PAR-2 expression was also significantly increased by A. fumigatus, which was significantly inhibited by FSLLRY-NH2. FSLLRY-NH2 significantly inhibited proinflammatory cytokine protein expression, as compared with that in infected control cells, which may be modified by p-ERK1/2. These data provide evidence that A. fumigatus increased PAR-2 expression and elevated disease, PMN infiltration, and proinflammatory cytokine expression through PAR-2, which may be modified by p-ERK1/2.

A structural model for the in vivo human cornea including collagen-swelling interaction

PubMed Central

Cheng, Xi; Petsche, Steven J.; Pinsky, Peter M.

2015-01-01

A structural model of the in vivo cornea, which accounts for tissue swelling behaviour, for the three-dimensional organization of stromal fibres and for collagen-swelling interaction, is proposed. Modelled as a binary electrolyte gel in thermodynamic equilibrium, the stromal electrostatic free energy is based on the mean-field approximation. To account for active endothelial ionic transport in the in vivo cornea, which modulates osmotic pressure and hydration, stromal mobile ions are shown to satisfy a modified Boltzmann distribution. The elasticity of the stromal collagen network is modelled based on three-dimensional collagen orientation probability distributions for every point in the stroma obtained by synthesizing X-ray diffraction data for azimuthal angle distributions and second harmonic-generated image processing for inclination angle distributions. The model is implemented in a finite-element framework and employed to predict free and confined swelling of stroma in an ionic bath. For the in vivo cornea, the model is used to predict corneal swelling due to increasing intraocular pressure (IOP) and is adapted to model swelling in Fuchs' corneal dystrophy. The biomechanical response of the in vivo cornea to a typical LASIK surgery for myopia is analysed, including tissue fluid pressure and swelling responses. The model provides a new interpretation of the corneal active hydration control (pump-leak) mechanism based on osmotic pressure modulation. The results also illustrate the structural necessity of fibre inclination in stabilizing the corneal refractive surface with respect to changes in tissue hydration and IOP. PMID:26156299

Diffusion of protein through the human cornea.

PubMed

Charalel, Resmi A; Engberg, Kristin; Noolandi, Jaan; Cochran, Jennifer R; Frank, Curtis; Ta, Christopher N

2012-01-01

To determine the rate of diffusion of myoglobin and bovine serum albumin (BSA) through the human cornea. These small proteins have hydrodynamic diameters of approximately 4.4 and 7.2 nm, and molecular weights of 16.7 and 66 kDa, for myoglobin and BSA, respectively. Diffusion coefficients were measured using a diffusion chamber where the protein of interest and balanced salt solution were in different chambers separated by an ex vivo human cornea. Protein concentrations in the balanced salt solution chamber were measured over time. Diffusion coefficients were calculated using equations derived from Fick's law and conservation of mass in a closed system. Our experiments demonstrate that the diffusion coefficient of myoglobin is 5.5 ± 0.9 × 10(-8) cm(2)/s (n = 8; SD = 1.3 × 10(-8) cm(2)/s; 95% CI: 4.6 × 10(-8) to 6.4 × 10(-8) cm(2)/s) and the diffusion coefficient of BSA is 3.1 ± 1.0 × 10(-8) cm(2)/s (n = 8; SD = 1.4 × 10(-8) cm(2)/s; 95% CI: 2.1 × 10(-8) to 4.1 × 10(-8) cm(2)/s). Our study suggests that molecules as large as 7.2 nm may be able to passively diffuse through the human cornea. With applications in pharmacotherapy and the development of an artificial cornea, further experiments are warranted to fully understand the limits of human corneal diffusion and its clinical relevance. Copyright © 2012 S. Karger AG, Basel.

Aniridia-related keratopathy: Structural changes in naïve and transplanted corneal buttons

PubMed Central

Stenevi, Ulf; Pedrosa Domellöf, Fátima

2018-01-01

Background To study structural changes in naïve and surgically treated corneas of aniridia patients with advanced aniridia-related keratopathy (ARK). Methods and findings Two naïve corneal buttons from patients with advanced ARK submitted to penetrating keratoplasty for the first time, one corneal button from an ARK patient that had undergone a keratolimbal allograft (KLAL), two corneal buttons from ARK patients who had previously undergone centered or decentered transplantation and were now retransplanted and two adult healthy donor control corneas were processed for immunohistochemistry. Antibodies against extracellular matrix components in the stroma and in the epithelial basement membrane (collagen I and IV, collagen receptor α11 integrin and laminin α3 chain), markers of fibrosis, wound healing and vascularization (fibronectin, tenascin-C, vimentin, α-SMA and caveolin-1), cell division (Ki-67) and macrophages (CD68) were used. Naïve ARK, KLAL ARK corneas and transplanted corneal buttons presented similar histopathological changes with irregular epithelium and disruption or absence of epithelial basal membrane. There was a loss of the orderly pattern of collagen lamellae and absence of collagen I in all ARK corneas. Vascularization was revealed by the presence of caveolin-1 and collagen IV in the pannus of all ARK aniridia corneas. The changes observed in decentered and centered transplants were analogous. Conclusions Given the similar pathological features of all cases, conditions inherent to the host seem to play an important role on the pathophysiology of the ARK in the long run. PMID:29889891

Aniridia-related keratopathy: Structural changes in naïve and transplanted corneal buttons.

PubMed

Vicente, André; Byström, Berit; Lindström, Mona; Stenevi, Ulf; Pedrosa Domellöf, Fátima

2018-01-01

To study structural changes in naïve and surgically treated corneas of aniridia patients with advanced aniridia-related keratopathy (ARK). Two naïve corneal buttons from patients with advanced ARK submitted to penetrating keratoplasty for the first time, one corneal button from an ARK patient that had undergone a keratolimbal allograft (KLAL), two corneal buttons from ARK patients who had previously undergone centered or decentered transplantation and were now retransplanted and two adult healthy donor control corneas were processed for immunohistochemistry. Antibodies against extracellular matrix components in the stroma and in the epithelial basement membrane (collagen I and IV, collagen receptor α11 integrin and laminin α3 chain), markers of fibrosis, wound healing and vascularization (fibronectin, tenascin-C, vimentin, α-SMA and caveolin-1), cell division (Ki-67) and macrophages (CD68) were used. Naïve ARK, KLAL ARK corneas and transplanted corneal buttons presented similar histopathological changes with irregular epithelium and disruption or absence of epithelial basal membrane. There was a loss of the orderly pattern of collagen lamellae and absence of collagen I in all ARK corneas. Vascularization was revealed by the presence of caveolin-1 and collagen IV in the pannus of all ARK aniridia corneas. The changes observed in decentered and centered transplants were analogous. Given the similar pathological features of all cases, conditions inherent to the host seem to play an important role on the pathophysiology of the ARK in the long run.

The influence of donor factors on corneal organ culture contamination.

PubMed

Gruenert, Anja K; Rosenbaum, Katja; Geerling, Gerd; Fuchsluger, Thomas A

2017-11-01

To evaluate the contamination rate and the corresponding spectrum of microbes and to identify donor risk factors for corneal organ culture contaminations. A total of 3306 organ-cultured donor corneas were included in the study. We performed a retrospective database analysis to evaluate donor factors such as gender, age, death-to-explantation interval (DEI), procurement site and cause of death and to determine their influence on donor cornea contaminations. Odds ratios (ORs) were calculated for each factor. The overall contamination rate was 7.8% (n = 259). Younger donor age (OR: 2.2, p = 0.003, chi-squared test), a DEI of more than 24 hr (OR: 1.6, p < 0.001), hospitalization prior to death (OR: 2.2, p < 0.001) and death caused by sepsis (OR: 2.7, p < 0.001) were associated with an increased risk of contamination, whereas donor gender did not have an effect on donor cornea contaminations. The most frequently isolated microbes were Enterococci (19%), Staphylococci (10.8%) and Candida (37.4%). This study helps to estimate the contamination risk of a cultured cornea based on specific donor factors. However, donors with risk factors should not be generally excluded from cornea donation. Further studies including antibiograms might clarify whether a change in the antibiotic composition of the culture medium would be useful to deal with the increasing number of multi-resistant microbes. © 2017 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

The Enigmatic Cornea and Intraocular Lens Calculations: The LXXIII Edward Jackson Memorial Lecture.

PubMed

Koch, Douglas D

2016-11-01

To review the progress and challenges in obtaining accurate corneal power measurements for intraocular lens (IOL) calculations. Personal perspective, review of literature, case presentations, and personal data. Through literature review findings, case presentations, and data from the author's center, the types of corneal measurement errors that can occur in IOL calculation are categorized and described, along with discussion of future options to improve accuracy. Advances in IOL calculation technology and formulas have greatly increased the accuracy of IOL calculations. Recent reports suggest that over 90% of normal eyes implanted with IOLs may achieve accuracy to within 0.5 diopter (D) of the refractive target. Though errors in estimation of corneal power can cause IOL calculation errors in eyes with normal corneas, greater difficulties in measuring corneal power are encountered in eyes with diseased, scarred, and postsurgical corneas. For these corneas, problematic issues are quantifying anterior corneal power and measuring posterior corneal power and astigmatism. Results in these eyes are improving, but 2 examples illustrate current limitations: (1) spherical accuracy within 0.5 D is achieved in only 70% of eyes with post-refractive surgery corneas, and (2) astigmatism accuracy within 0.5 D is achieved in only 80% of eyes implanted with toric IOLs. Corneal power measurements are a major source of error in IOL calculations. New corneal imaging technology and IOL calculation formulas have improved outcomes and hold the promise of ongoing progress. Copyright © 2016 Elsevier Inc. All rights reserved.

Defining a mechanistic link between pigment epithelium-derived factor, docosahexaenoic acid, and corneal nerve regeneration.

PubMed

Pham, Thang Luong; He, Jiucheng; Kakazu, Azucena H; Jun, Bokkyoo; Bazan, Nicolas G; Bazan, Haydee E P

2017-11-10

The cornea is densely innervated to sustain the integrity of the ocular surface. Corneal nerve damage produced by aging, diabetes, refractive surgeries, and viral or bacterial infections impairs tear production, the blinking reflex, and epithelial wound healing, resulting in loss of transparency and vision. A combination of the known neuroprotective molecule, pigment epithelium-derived factor (PEDF) plus docosahexaenoic acid (DHA), has been shown to stimulate corneal nerve regeneration, but the mechanisms involved are unclear. Here, we sought to define the molecular events of this effect in an in vivo mouse injury model. We first confirmed that PEDF + DHA increased nerve regeneration in the mouse cornea. Treatment with PEDF activates the phospholipase A 2 activity of the PEDF-receptor (PEDF-R) leading to the release of DHA; this free DHA led to enhanced docosanoid synthesis and induction of bdnf, ngf , and the axon growth promoter semaphorin 7a ( sema7a ), and as a consequence, their products appeared in the mouse tears. Surprisingly, corneal injury and treatment with PEDF + DHA induced transcription of neuropeptide y ( npy ), small proline-rich protein 1a ( sprr1a ), and vasoactive intestinal peptide ( vip ) in the trigeminal ganglia (TG). The PEDF-R inhibitor, atglistatin, blocked all of these changes in the cornea and TG. In conclusion, we uncovered here an active cornea-TG axis, driven by PEDF-R activation, that fosters axon outgrowth in the cornea. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

West Nile Virus Infection in Human and Mouse Cornea Tissue

PubMed Central

Blitvich, Bradley J.; Wang, Tian; Saxena, Vandana; Zeng, Shemin; Harmon, Karen M.; Raymond, Matthew D.; Goins, Kenneth M.; Reed, Cynthia R.; Mullins, Robert F.; Greiner, Mark A.

2016-01-01

The purpose of this study was to determine the in vitro and ex vivo susceptibility of human corneal cells to West Nile virus (WNV) infection and evaluate the ability of the virus to disseminate to the corneas of infected mice. Human corneal epithelial cells were challenged with WNV, incubated for 1–6 days, and tested for evidence of WNV infection. Viral RNA and antigen were detected at every time point, and the virus reached a peak titer of 2.5 × 107 plaque-forming units (pfu)/mL at 3 days postinoculation (PI). Corneas procured from donors were incubated in culture dishes containing WNV for 1–5 days and tested for evidence of WNV. Viral RNA and antigen were detected, and the virus reached a mean peak titer of 4.9 × 104 pfu/mL at 5 days PI. Mice were inoculated intraperitoneally with WNV, and their eyes were harvested at 2, 5, and 8 days PI and tested for evidence of WNV. Viral RNA was detected in corneas of four of nine systemically infected mice as early as 2 days PI. We conclude that human corneal cells support WNV replication in vitro and ex vivo, and WNV may disseminate into the corneas of experimentally infected mice. These findings indicate that corneal transmission cannot be ruled out as a novel mode of human-to-human WNV transmission and additional experiments should be conducted to assess this risk further. PMID:27672204

Mitochondrial DNA common deletion in the human eye: a relation with corneal aging.

PubMed

Gendron, Sébastien P; Mallet, Justin D; Bastien, Nathalie; Rochette, Patrick J

2012-01-01

The most frequent mitochondrial DNA (mtDNA) mutation is a 4977 bp deletion known as the common deletion (mtDNA(CD4977)). mtDNA(CD4977) is related to skin photo-aging and to chronological aging of cells with high-energy demands such as neurons and muscle cells. The human eye contains both sun-exposed (cornea, iris) and high-energy demand structures (retina). In this study, we employed a highly sensitive quantitative PCR technique to determine mtDNA(CD4977) occurrence in different structures of the human eye. We found that the cornea, the most anterior structure of the eye, contains the highest amount of mtDNA(CD4977) (2.6%, 0.25% and 0.06% for the cornea, iris and retina, respectively). Within the cornea, mtDNA(CD4977) is almost exclusively found in the stroma, the cellular layer conferring transparency and rigidity to the human cornea (8.59%, 0.13% and 0.05% in the stroma, endothelium and epithelium, respectively). Moreover, we show that mtDNA(CD4977) accumulates with age in the corneal stroma. Taken together, our results suggest that mtDNA(CD4977) is related to photo-aging rather than chronological aging in the human eye. Similar to the involvement of mtDNA(CD4977) in skin photo-aging phenotypes, we believe that the clinical manifestations of corneal aging, including clouding and stiffening, are associated with the accumulation of mtDNA(CD4977) in the corneal stroma. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

The Influence of Lamellar Orientation on Corneal Material Behavior: Biomechanical and Structural Changes in an Avian Corneal Disorder

PubMed Central

Elsheikh, Ahmed; Kassem, Wael; Kamma-Lorger, Christina S.; Hocking, Paul M.; White, Nick; Inglehearn, Chris F.; Ali, Manir; Meek, Keith M.

2011-01-01

Purpose. Retinopathy, globe enlarged (RGE) is an inherited genetic disease of chickens with a corneal phenotype characterized by loss of tissue curvature and changes in peripheral collagen fibril alignment. This study aimed to characterize the material behavior of normal and RGE chicken corneas under inflation and compare this with new spatial- and depth-resolved microstructural information to investigate how stromal fibril architecture determines corneal behavior under intraocular pressure (IOP). Methods. Six RGE chicken corneas and six age-matched normal controls were tested using trephinate inflation and their stress-strain behavior determined as a function of posterior pressure. Second harmonic generation mulitphoton microscopy was used to compare the in-plane appearance and degree of through-plane interlacing of collagen lamellae between normal and mutant corneas. Results. RGE corneas displayed a 30–130% increase in material stiffness [Etangent(RGE) = 0.94 ± 0.18 MPa to 3.09 ± 0.66 MPa; Etangent(normals) = 0.72 ± 0.13 MPa to 1.34 ± 0.35 MPa] (P ≤ 0.05). The normal in-plane disposition of anterior collagen in the peripheral cornea was altered in RGE but through-plane lamellar interlacing was unaffected. Conclusions. This article demonstrates changes in corneal material behavior in RGE that are qualitatively consistent with microstructural collagen alterations identified both herein and previously. This study indicates that, in general, changes in stromal fibril orientation may significantly affect corneal material behavior and thereby its response to IOP. PMID:21051696

Promoting effect of borneol on the permeability of puerarin eye drops and timolol maleate eye drops through the cornea in vitro.

PubMed

Wu, Chun-jie; Huang, Qin-wan; Qi, Hong-yi; Guo, Ping; Hou, Shi-xiang

2006-09-01

Studies on the influence of borneol on the penetration of puerarin eye drops and timolol maleate eye drops through the cornea, and evaluation of the ocular irritability were conducted to provide a theoretical basis for the application of borneol in enhancing corneal permeability. The cornea penetrative experiment in vitro was conducted to observe the quantitative change of puerarin and timolol maleate penetrated through the cornea after administering different dosages of borneol. The corneal hydration level and blinking frequency were recorded as irritability indexes in vitro and in vivo. The steady-flow J of high, middle and low dosage groups of puerarin eye drops with borneol were increased by 49%, 32%, 5% respectively, and permeability parameter Kp increased by 49%, 32%, 5% respectively, as compared to that of the control group. The steady-flow J of high dosage group of timolol maleate eye drops with borneol was increased by 5%; middle and low dosage groups with borneol were decreased by 6%, 3% respectively. The permeability parameter Kp of high dosage group increased by 5%, while middle and low dosage groups with borneol were decreased by 6%, 3% respectively, as compared to that of the control group. Evaluation showed no ocular irritability caused by borneol. The results of this study suggest that the promoting effect of borneol on the permeability of drugs through the cornea in vitro is selective, which indicates that borneol has the potential to be used as an ophthalmic penetration enhancer.

West Nile Virus Infection in Human and Mouse Cornea Tissue.

PubMed

Blitvich, Bradley J; Wang, Tian; Saxena, Vandana; Zeng, Shemin; Harmon, Karen M; Raymond, Matthew D; Goins, Kenneth M; Reed, Cynthia R; Mullins, Robert F; Greiner, Mark A

2016-11-02

The purpose of this study was to determine the in vitro and ex vivo susceptibility of human corneal cells to West Nile virus (WNV) infection and evaluate the ability of the virus to disseminate to the corneas of infected mice. Human corneal epithelial cells were challenged with WNV, incubated for 1-6 days, and tested for evidence of WNV infection. Viral RNA and antigen were detected at every time point, and the virus reached a peak titer of 2.5 × 10 7 plaque-forming units (pfu)/mL at 3 days postinoculation (PI). Corneas procured from donors were incubated in culture dishes containing WNV for 1-5 days and tested for evidence of WNV. Viral RNA and antigen were detected, and the virus reached a mean peak titer of 4.9 × 10 4 pfu/mL at 5 days PI. Mice were inoculated intraperitoneally with WNV, and their eyes were harvested at 2, 5, and 8 days PI and tested for evidence of WNV. Viral RNA was detected in corneas of four of nine systemically infected mice as early as 2 days PI. We conclude that human corneal cells support WNV replication in vitro and ex vivo, and WNV may disseminate into the corneas of experimentally infected mice. These findings indicate that corneal transmission cannot be ruled out as a novel mode of human-to-human WNV transmission and additional experiments should be conducted to assess this risk further. © The American Society of Tropical Medicine and Hygiene.

3D refraction correction and extraction of clinical parameters from spectral domain optical coherence tomography of the cornea.

PubMed

Zhao, Mingtao; Kuo, Anthony N; Izatt, Joseph A

2010-04-26

Capable of three-dimensional imaging of the cornea with micrometer-scale resolution, spectral domain-optical coherence tomography (SDOCT) offers potential advantages over Placido ring and Scheimpflug photography based systems for accurate extraction of quantitative keratometric parameters. In this work, an SDOCT scanning protocol and motion correction algorithm were implemented to minimize the effects of patient motion during data acquisition. Procedures are described for correction of image data artifacts resulting from 3D refraction of SDOCT light in the cornea and from non-idealities of the scanning system geometry performed as a pre-requisite for accurate parameter extraction. Zernike polynomial 3D reconstruction and a recursive half searching algorithm (RHSA) were implemented to extract clinical keratometric parameters including anterior and posterior radii of curvature, central cornea optical power, central corneal thickness, and thickness maps of the cornea. Accuracy and repeatability of the extracted parameters obtained using a commercial 859nm SDOCT retinal imaging system with a corneal adapter were assessed using a rigid gas permeable (RGP) contact lens as a phantom target. Extraction of these parameters was performed in vivo in 3 patients and compared to commercial Placido topography and Scheimpflug photography systems. The repeatability of SDOCT central corneal power measured in vivo was 0.18 Diopters, and the difference observed between the systems averaged 0.1 Diopters between SDOCT and Scheimpflug photography, and 0.6 Diopters between SDOCT and Placido topography.

Corneal Tissue From Dry Eye Donors Leads to Enhanced Graft Rejection.

PubMed

Inomata, Takenori; Hua, Jing; Nakao, Takeshi; Shiang, Tina; Chiang, Homer; Amouzegar, Afsaneh; Dana, Reza

2018-01-01

To assess the effect of dry eye disease (DED) in graft donors on dendritic cell (DC) maturation, host T-cell sensitization, and corneal allograft rejection. Corneas of control (healthy donor) and DED mice (C57BL/6) were transplanted onto fully allogeneic naive BALB/c recipients (n = 10 mice/group). Long-term allograft survival was evaluated for 8 weeks. Corneas and draining lymph nodes (dLNs) were harvested at posttransplantation day 14 (n = 5 mice/group). The frequencies of MHCII CD11c DCs in the donor corneas and host dLNs and the frequencies of interferon (IFN)-γ and IL-17 CD4 T cells and Foxp3 expression by Tregs in host dLNs were investigated using flow cytometry. The enzyme-linked immunospot assay was used to assess host T-cell allosensitization through direct and indirect pathways (n = 3/group). Recipients of DED donor corneas showed significantly reduced graft survival (10%) compared with control mice (50% survival, P = 0.022), and had significantly increased frequencies of mature DCs in the grafted cornea (DED donor 44.0% ± 0.36% vs. healthy donor 35.4 ± 0.5%; P < 0.0001) and host dLNs (DED donor 25.1% ± 0.66% vs. healthy donor 13.7% ± 1.6%; P = 0.005). Frequencies of IFN-γ and IL-17 T cells were increased in the dLNs of recipients of DED corneas, whereas the expression (mean fluorescence intensity) of Foxp3 in Tregs was decreased significantly in these mice (DED donor 6004 ± 193 vs. healthy donor 6806 ± 81; P = 0.0002). Enzyme-linked immunospot analysis showed that the direct pathway of allosensitization was significantly amplified in recipients of grafts with DED (P = 0.0146). Our results indicate that DED in the donor is a significant risk factor for subsequent corneal allograft rejection.

Cornea Preservation Time Study: Methods and Potential Impact on the Cornea Donor Pool in the United States

PubMed Central

Szczotka-Flynn, Loretta B.; Ayala, Allison R.; Benetz, Beth A.; Gal, Robin L.; Aldave, Anthony J.; Corrigan, Michelle M.; Dunn, Steven P.; McCall, Ty L.; Pramanik, Sudeep; Rosenwasser, George O.; Ross, Kevin W.; Terry, Mark A.; Verdier, David D.

2015-01-01

Purpose: The aim of this study was to describe the aims, methods, donor and recipient cohort characteristics, and potential impact of the Cornea Preservation Time Study (CPTS). Methods: The CPTS is a randomized clinical trial conducted at 40 clinical sites (70 surgeons) designed to assess the effect of donor cornea preservation time (PT) on graft survival 3 years after Descemet stripping automated endothelial keratoplasty (DSAEK). Eyes undergoing surgery for Fuchs endothelial corneal dystrophy or pseudophakic/aphakic corneal edema were randomized to receive donor corneas stored ≤7 days or 8 to 14 days. Donor and patient characteristics, tissue preparation and surgical parameters, recipient and donor corneal stroma clarity, central corneal thickness, intraocular pressure, complications, and a reading center-determined central endothelial cell density were collected. Surveys were conducted to evaluate pre-CPTS PT practices. Results: The 1330 CPTS donors were: 49% >60 years old, 27% diabetic, had a median eye bank–determined screening endothelial cell density of 2688 cells/mm2, and 74% eye bank prepared for DSAEK. A total of 1090 recipients (1330 eyes including 240 bilateral cases) had: median age of 70 years, were 60% female, 90% white, 18% diabetic, 52% phakic, and 94% had Fuchs endothelial corneal dystrophy. Before the CPTS, 19 eye banks provided PT data on 20,852 corneas domestically placed for DSAEK in 2010 to 2011; 96% were preserved ≤7 days. Of 305 American Academy of Ophthalmology members responding to a pre-CPTS survey, 233 (76%) set their maximum PT preference at 8 days or less. Conclusions: The CPTS will increase understanding of factors related to DSAEK success and, if noninferiority of longer PT is shown, will have great potential to extend the available pool of endothelial keratoplasty donors. Clinical Trial Registration—URL: http://www.clinicaltrials.gov. Unique identifier: NCT01537393. PMID:25850706

A non-surgical approach to the management of exposure keratitis due to facial palsy by using mini-scleral lenses.

PubMed

Zaki, Victor

2017-02-01

This is a retrospective study aimed to determine the efficacy of mini-scleral contact lens in protecting the cornea and improving vision in cases of facial palsy. Patients with facial palsy get exposure keratitis because the cornea is dry. They feel pain, discomfort and excessive watering. If left untreated, it leads to permanent damage to the cornea and loss of good functional vision. Mini-scleral lens keep the cornea covered by saline solution all wearing hours. Three patients (4 eyes) with acoustic neuroma, two unilateral and one bilateral, who underwent acoustic neuroma surgeries resulting in facial palsy, are presented.The gold implant and lateral tarsorrhaphy were not enough for corneal protection.Two patients (patients 1 and 2) suffered continuous pain and watering. They had to apply thick lubricant, Lacri-Lube ointment (Allergan, Inc., Dublin, Ireland), several times daily to the affected eye for 15 years. The vision of these patients in the affected eyes were counting fingers (CF) at one foot.Patient 3 with bilateral facial palsy had exposure keratitis in both eyes resulting in constant watering, pain and blurred vision. The 4 eyes were fitted with mini-scleral lenses. The lenses were 15.8 mm rigid gas permeable filled with preservative free saline solution that continuously covers the cornea all wearing hours. In patients 1 and 2 with unilateral facial palsy, vision improved through the mini-scleral lenses to 20/30 and all their symptoms disappeared.The keratitis in case 3 with bilateral facial palsy disappeared within one week of mini-scleral lens use.Follow up for 2 years showed that these patients maintained good vision with no side effects. Mini-scleral lenses protected the cornea, gave comfort and improved the vision and the quality of life of these three patients with facial palsy and should be considered for all patients with facial palsy.

The effect of single and repeated UVB radiation on rabbit cornea.

PubMed

Fris, Miroslav; Tessem, May-Britt; Cejková, Jitka; Midelfart, Anna

2006-12-01

Cumulative effect of ultraviolet radiation (UVR) is an important aspect of UV corneal damage. The purpose of this study was to apply high resolution magic angle spinning proton nuclear magnetic resonance (HR-MAS 1H NMR) spectroscopy to evaluate the effect of single and repeated UV radiation exposure of the same overall dose on the rabbit cornea. Corneal surfaces of 24 normal rabbit eyes were examined for the effects of UVB exposure (312 nm). In the first group (UVB1), animals were irradiated with a single dose (3.12 J/cm2; 21 min) of UVB radiation. The animals in the second group (UVB2) were irradiated three times for 7 min every other day (dose of 1.04 J/cm2; days 1, 3, 5) to give the same overall dose (3.12 J/cm2). The third group served as an untreated control group. One day after the last irradiation, the animals were sacrificed, and the corneas were removed and frozen. HR-MAS 1H NMR spectra from intact corneas were obtained. Special grouping patterns among the tissue samples and the relative percentage changes in particular metabolite concentrations were evaluated using modern statistical methods (multivariate analysis, one-way ANOVA). The metabolic profile of both groups of UVB-irradiated samples was significantly different from the control corneas. Substantial decreases in taurine, hypo-taurine and choline-derivatives concentrations and substantial elevation in glucose and betaine levels were observed following the UVR exposure. There was no significant difference between the effect of a single and repeated UVB irradiation of the same overall dose. For the first time, the effects of single and repeated UVR doses on the metabolic profile of the rabbit cornea were analysed and compared. The combination of HR-MAS 1H NMR spectroscopy and modern statistical methods (multivariate analysis, one-way ANOVA) proved suitable to assess the overall view of the metabolic alterations in the rabbit corneal tissue following UVB radiation exposure.

Interface fluid syndrome in human eye bank corneas after LASIK: causes and pathogenesis.

PubMed

Dawson, Daniel G; Schmack, Ingo; Holley, Glenn P; Waring, George O; Grossniklaus, Hans E; Edelhauser, Henry F

2007-10-01

To evaluate the effects of corneal edema on human donor corneas that had previous LASIK using a laboratory model with histologic and ultrastructural correlations. Experimental study. Thirty human eye bank corneas from 15 donors (mean age +/- standard deviation, 49.9+/-8.9 years) who had had previous LASIK surgery (2-8 years before death). The corneas were mounted in an artificial anterior chamber and the corneal endothelium was perfused for up to 5.0 hours with 0.9% saline solution (endothelial cell damage group) or BSS Plus at a pressure of 15 mmHg (control group), or BSS Plus at a pressure of 55 mmHg (high-pressure group). The corneas were evaluated by confocal and specular microscopy before, during, and at the end of the experimental period. Subsequently, the specimens were evaluated by light and electron microscopy. Corneal thickness, reflectivity, histology, and ultrastructure. Endothelial cell damage resulted in an increased (141.5+/-38.8 microm) total corneal thickness relative to controls (52.3+/-33.7 microm), whereas high pressure resulted in a decreased thickness (24.8+/-14.1 microm) relative to controls. This ultimately was due to swelling of the LASIK interface in both groups and swelling of the residual stromal bed (RSB) in the endothelial cell damage group or compression of the RSB and, possibly, the flap in the high-pressure group. A significant increase in corneal reflectivity at the LASIK interface occurred in both groups, primarily due to varying degrees of fluid accumulation and associated hydropic keratocyte degeneration, as well as increased corneal reflectivity in the RSB only in the endothelial cell damage group. After LASIK surgery, edematous corneas preferentially hydrate and swell in the paracentral and central interface wound, commonly resulting in a hazy corneal appearance primarily due to keratocyte hydropic degeneration. More severe corneal edema is characterized by the formation of an optically empty space corresponding to an interface fluid pocket. The spectrum of interface fluid syndrome can be described in 3 stages.

Evaluation of Biomechanical Changes in Myopia Patients with Unsatisfactory Corneas After Femto Second-Laser In Situ Keratomileusis (FS-LASIK) Concurrent with Accelerated Corneal Collagen Cross-Linking Using Corvis-ST: Two-Year Follow-Up Results.

PubMed

Xu, Weiwei; Tao, Ye; Wang, Liqiang; Huang, Yifei

2017-07-27

BACKGROUND Some myopia patients with unsatisfactory corneas consider corneal refractive surgery for different reasons. Accelerated corneal collagen crosslinking (ACXL) is an effective method to enhance the resistance of the cornea. The present investigation was designed to evaluate the changes of biomechanical properties in patients with myopia and thin corneas after femtosecond-laser in situ keratomileusis (FS-LASIK) concurrent with ACXL. MATERIAL AND METHODS A prospective study was designed. A total of 22 eyes of 11 myopia astigmatism patients with unsatisfactory corneas were enrolled. The patients were assigned to femtosecond-laser in situ keratomileusis concurrent with accelerated corneal collagen crosslinking (FS-LASIK-ACXL). The follow-up duration was 24 months. Manifest refraction, uncorrected (UDVA), and corrected distance visual acuity (CDVA), ultra-high-speed camera (Corvis-ST), corneal topography, anterior segment OCT (AS-OCT), Pentacam, and endothelial cell density (ECD) were examined before and after the operation. The corneal biomechanical and refractive data was analyzed using SAS9.3. Data were analyzed through normal distribution test and variance of analysis. The difference was considered as statistically significant when p<0.05. RESULTS The steep K (Ks) values, flat K (Kf) values, average keratometry values (Avek) values, and central corneal thickness (CCT) declined significantly after FS-LASIK-ACXL operation. The values of first applanation length (A1L), the second applanation length (A2L), the first applanation velocity (A1V), the second applanation velocity (A2V), deformation amplitude (DA), highest concavity peak distance (PD), and radius of curvature at the time of highest concavity (HCR) did not show significant difference after the operation. CONCLUSIONS FS-LASIK-ACXL is an effective and safe surgery for improving visual acuity for myopic patients with thin corneas, and it does not increase the risk of iatrogenic keratectasia.

EXPRESSION OF NeuGc ON PIG CORNEAS AND ITS POTENTIAL SIGNIFICANCE IN PIG CORNEAL XENOTRANSPLANTATION

PubMed Central

Lee, Whayoung; Miyagawa, Yuko; Long, Cassandra; Ekser, Burcin; Walters, Eric; Ramsoondar, Jagdeece; Ayares, David; Tector, A. Joseph; Cooper, David K. C.; Hara, Hidetaka

2016-01-01

Purpose Pigs expressing neither galactose-α1,3-galactose (Gal) nor N-glycolylneuraminic acid (NeuGc) take xenotransplantation one step closer to the clinic. Our aims were (i) to document the lack of NeuGc expression on corneas and aortas, and cultured endothelial cells (aortic [AECs]; corneal [CECs]) of GTKO/NeuGcKO pigs, and (ii) to investigate whether the absence of NeuGc reduced human antibody binding to the tissues and cells. Methods Wild-type (WT), GTKO, and GTKO/NeuGcKO pig were used for the study. Human tissues and cultured cells were negative controls. Immunofluorescence staining was performed using anti-Gal and anti-NeuGc antibodies, and to determine human IgM and IgG binding to tissues. Flow cytometric analysis was used to determine Gal and NeuGc expression on cultured CECs and AECs and to measure human IgM/IgG binding to these cells. Results Both Gal and NeuGc were detected on WT pig corneas and aortas. Although GTKO pigs expressed NeuGc, neither human nor GTKO/NeuGcKO pigs expressed Gal or NeuGc. Human IgM/IgG binding to corneas and aortas from GTKO and GTKO/NeuGcKO pigs was reduced compared to binding to WT pigs. Human antibody binding to GTKO/NeuGcKO AECs was significantly less than to GTKO AECs, but there was no significant difference in binding between GTKO and GTKO/NeuGcKO CECs. Conclusions The absence of NeuGc on GTKO aortic tissue and AECs is associated with reduced human antibody binding, and possibly will provide better outcome in clinical xenotransplantation using vascularized organs. For clinical corneal xenotransplantation, the absence of NeuGc expression on GTKO/NeuGcKO pig corneas may not prove an advantage over GTKO corneas. PMID:26418433

Molecular Evidence and Functional Expression of a Novel Drug Efflux pump (ABCC2) in Human Corneal Epithelium and Rabbit Cornea and its role in Ocular drug efflux

PubMed Central

Karla, Pradeep K.; Pal, Dhananjay; Quinn, Tim; Mitra, Ashim K.

2007-01-01

Cornea is considered as a major barrier for ocular drug delivery. Low ocular bioavailability of drugs has been attributed primarily to low permeability across corneal epithelium thus leading to sub-therapeutic concentrations of drug in the eye and treatment failure. The role of drug efflux proteins, particularly the Pglycoprotein in ocular drug bioavailability has been reported. The objective of this research was to determine whether human corneal epithelium expresses multi drug resistance associated proteins contributing to drug efflux by employing both cultured corneal cells and freshly excised rabbit cornea. SV40 HCEC and rPCEC were selected for in-vitro testing. SV40-HCEC and freshly excised rabbit corneas were utilized for transport studies. [3H]-cyclosporine-A and [14C]-erythromycin which are known substrates for ABCC2 and MK-571, a specific inhibitor for MRP were applied in this study. RT-PCR indicated a unique and distinct band at ∼272 bp corresponding to ABCC2 in HCEC, SV40-HCEC, rabbit cornea, rPCEC and MDCKII-MRP2 cells. Also RT-PCR indicated a unique band ∼181 bp for HCEC and SV40-HCEC. Immunoprecipitation followed by Western Blot analysis revealed a specific band at ∼190-kDa in membrane fraction of SV40-HCEC, MDCKII-MRP2 and no band with isotype control. Uptake of [3H]-cyclosporine-A and [14C]-erythromycin in the presence of MK-571 was significantly enhanced than control in both SV40 HCEC and rPCEC. Similarly a significant elevation in (A→B) permeability of [3H]-cyclosporine-A and [14C]-erythromycin was observed in the presence of MK-571 in SV40-HCEC. A→B transport of [3H]-cyclosporine-A was elevated in the presence of MK-571 in freshly excised rabbit cornea indicating potential role of this efflux transporter and high clinical significance of this finding. PMID:17156953

Corneal Expression of SLURP-1 by Age, Sex, Genetic Strain, and Ocular Surface Health

PubMed Central

Swamynathan, Sudha; Delp, Emili E.; Harvey, Stephen A. K.; Loughner, Chelsea L.; Raju, Leela; Swamynathan, Shivalingappa K.

2015-01-01

Purpose Although secreted Ly6/urokinase-type plasminogen activator receptor–related protein-1 (Slurp1) transcript is highly abundant in the mouse cornea, corresponding protein expression remains uncharacterized. Also, SLURP1 was undetected in previous tear proteomics studies, resulting in ambiguity about its baseline levels. Here, we examine mouse corneal Slurp1 expression in different sexes, age groups, strains, and health conditions, and quantify SLURP1 in human tears from healthy or inflamed ocular surfaces. Methods Expression of Slurp1 in embryonic day-13 (E13), E16, postnatal day-1 (PN1), PN10, PN20, and PN70 Balb/C, FVBN, C57Bl/6, and DBA/2J mouse corneas, Klf4Δ/ΔCE corneas with corneal epithelial–specific ablation of Klf4, migrating cells in wild-type corneal epithelial wound edge, and in corneas exposed to pathogen-associated molecular patterns (PAMPs) poly(I:C), zymosan-A, or Pam3Csk4 was examined by QPCR, immunoblots, and immunofluorescent staining. Human SLURP1 levels were quantified by ELISA in tears from 34 men and women aged 18 to 80 years. Results Expression of Slurp1, comparable in different strains and sexes, was low in E13, E16, PN1, and PN10 mouse corneas, and increased rapidly after eyelid opening in a Klf4-dependent manner. We found Slurp1 was downregulated in corneas exposed to PAMPs, and in migrating cells at the wound edge. Human SLURP1 expression, comparable in different sexes and age groups, was significantly decreased in tears from inflamed ocular surfaces (0.34%) than those from healthy individuals (0.77%). Conclusions These data describe the influence of age, sex, genetic background, and ocular surface health on mouse corneal expression of Slurp1, establish the baseline for human tear SLURP1 expression, and identify SLURP1 as a useful diagnostic and/or therapeutic target for inflammatory ocular surface disorders. PMID:26670825

Outcomes of descemet stripping automated endothelial keratoplasty using imported donor corneas.

PubMed

Lekhanont, Kaevalin; Vanikieti, Kavin; Nimvorapun, Nutthida; Chuckpaiwong, Varintorn

2017-04-05

The lack of development of local donor tissue acquisition in several regions of the world has resulted in the necessity of performing keratoplasty with imported donor corneas. The greatest concern about the use of donor corneas supplied by foreign eye banks is the effect of the increased donor death-to-operation time which inevitably occurs during the tissue recovery, tissue processing, and tissue transfer between the countries. The purpose of this study was to report the outcomes of descemet stripping automated endothelial keratoplasty (DSAEK) using imported donor corneas. This retrospective, non-comparative case series investigated the outcomes of the 102 consecutive DSAEK procedures using imported donor corneas performed at a single university-based hospital between August 2006-2014. The main outcome measures were postoperative best-corrected visual acuity (BCVA), endothelial cell density (ECD), and complications. The mean death-to-operation time was 9.52 ± 1.48 days (range, 8-13). The mean preoperative ECD was 2761 ± 285 cells/mm 2 . Fuchs' endothelial dystrophy was the predominant indication for grafting. The mean follow-up duration was 65.3 months. Ninety-three eyes had improved vision postoperatively (91.18%). BCVA unchanged in 3 eyes due to preexisting macular scar and advanced glaucoma. Primary graft failure occurred in 6 eyes (5.88%). Of the 93 eyes with improved BCVA, 100% had their best corrected vision within the first 1 year. The mean ECD at 6, 12, 24, 36, and 60 months after surgery was 1762 ± 294 cells/mm 2 , 1681 ± 284 cells/mm 2 , 1579 ± 209 cells/mm 2 , 1389 ± 273 cells/mm 2 , and 1251 ± 264 cells/mm 2 respectively. The mean ECD loss at 6 months, 1 year, 2 years, 3 years, and 5 years after surgery was 36.2%, 39.1%, 42.8%, 49.7%, and 54.7% respectively. The most common complication was graft detachment/dislocation (10.78%). There were no cases of any postoperative infection. DSAEK with imported donor corneas provides rapid and good visual rehabilitation. The percentages of endothelial cell loss were comparable to those achieved in Western series using domestic corneas in which fresher tissues were available for transplantation.

Correlation between epithelial thickness in normal corneas, untreated ectatic corneas, and ectatic corneas previously treated with CXL; is overall epithelial thickness a very early ectasia prognostic factor?

PubMed

Kanellopoulos, Anastasios John; Aslanides, Ioannis M; Asimellis, George

2012-01-01

To determine and correlate epithelial corneal thickness (pachymetric) measurements taken with a digital arc scanning very high frequency ultrasound biomicroscopy (HF UBM) imaging system (Artemis-II), and compare mean and central epithelial thickness among normal eyes, untreated keratoconic eyes, and keratoconic eyes previously treated with collagen crosslinking (CXL). Epithelial pachymetry measurements (topographic mapping) were conducted on 100 subjects via HF UBM. Three groups of patients were included: patients with normal eyes (controls), patients with untreated keratoconic eyes, and patients with keratoconic eyes treated with CXL. Central, mean, and peripheral corneal epithelial thickness was examined for each group, and a statistical study was conducted. Mean, central, and peripheral corneal epithelial thickness was compared between the three groups of patients. Epithelium thickness varied substantially in the keratoconic group, and in some cases there was a difference of up to 20 μm between various points of the same eye, and often a thinner epithelium coincided with a thinner cornea. However, on average, data from the keratoconic group suggested an overall thickening of the epithelium, particularly over the pupil center of the order of +3 μm, while the mean epithelium thickness was on average +1.1 μm, compared to the control population (P = 0.005). This overall thickening was more pronounced in younger patients in the keratoconic group. Keratoconic eyes previously treated with CXL showed, on average, virtually the same average epithelium thickness (mean -0.7 μm, -0.2 μm over the pupil center, -0.9 μm over the peripheral zone) as the control group. This finding further reinforces our novel theory of the "reactive" component of epithelial thickening in corneas that are biomechanically unstable, becoming stable when biomechanical rigidity is accomplished despite persistence of cornea topographic irregularity. A highly irregular epithelium may be suggestive of an ectatic cornea. Our results indicate that the epithelium is thinner over the keratoconic protrusion, but to a much lesser extent than anticipated, and on average epithelium is thicker in this group of patients. This difference appears to be clinically significant and may become a screening tool for eyes suspected for ectasia.

Correlation between epithelial thickness in normal corneas, untreated ectatic corneas, and ectatic corneas previously treated with CXL; is overall epithelial thickness a very early ectasia prognostic factor?

PubMed Central

Kanellopoulos, Anastasios John; Aslanides, Ioannis M; Asimellis, George

2012-01-01

Purpose To determine and correlate epithelial corneal thickness (pachymetric) measurements taken with a digital arc scanning very high frequency ultrasound biomicroscopy (HF UBM) imaging system (Artemis-II), and compare mean and central epithelial thickness among normal eyes, untreated keratoconic eyes, and keratoconic eyes previously treated with collagen crosslinking (CXL). Methods Epithelial pachymetry measurements (topographic mapping) were conducted on 100 subjects via HF UBM. Three groups of patients were included: patients with normal eyes (controls), patients with untreated keratoconic eyes, and patients with keratoconic eyes treated with CXL. Central, mean, and peripheral corneal epithelial thickness was examined for each group, and a statistical study was conducted. Results Mean, central, and peripheral corneal epithelial thickness was compared between the three groups of patients. Epithelium thickness varied substantially in the keratoconic group, and in some cases there was a difference of up to 20 μm between various points of the same eye, and often a thinner epithelium coincided with a thinner cornea. However, on average, data from the keratoconic group suggested an overall thickening of the epithelium, particularly over the pupil center of the order of +3 μm, while the mean epithelium thickness was on average +1.1 μm, compared to the control population (P = 0.005). This overall thickening was more pronounced in younger patients in the keratoconic group. Keratoconic eyes previously treated with CXL showed, on average, virtually the same average epithelium thickness (mean −0.7 μm, −0.2 μm over the pupil center, −0.9 μm over the peripheral zone) as the control group. This finding further reinforces our novel theory of the “reactive” component of epithelial thickening in corneas that are biomechanically unstable, becoming stable when biomechanical rigidity is accomplished despite persistence of cornea topographic irregularity. Conclusion A highly irregular epithelium may be suggestive of an ectatic cornea. Our results indicate that the epithelium is thinner over the keratoconic protrusion, but to a much lesser extent than anticipated, and on average epithelium is thicker in this group of patients. This difference appears to be clinically significant and may become a screening tool for eyes suspected for ectasia. PMID:22701079

Simulation of Blast Loading on an Ultrastructurally-based Computational Model of the Ocular Lens

DTIC Science & Technology

2012-10-01

mechanisms of traumatic cataract forma - tion that may require cataract surgery (implantation of an intraocular lens (IOL)) are not well understood in...cornea and cutting through the iris and zonules. This was done by making a small incision with a diamond-head blade at the edge of the cornea and using

The brilliant beauty of the eye: light reflex from the cornea and tear film.

PubMed

Goto, Eiki

2006-12-01

Light reflex from the cornea and tear film as contributors to beautiful eyes ("eye sparkling") are reviewed. A systematic literature review was conducted using "Purkinje-Sanson image," "corneal light reflex," "corneal topography," "corneal wavefront aberration," and "tear interference image" as search terms. Articles on corneal surface regularity and stability and tear interferometry of the precorneal tear lipid layer were reviewed. PS-1 image, that is light reflex from the cornea and tear film, is widely used in practical ophthalmic examination. To achieve a brilliant beauty of the eye ("eye sparkling"), it is important that the tear film (aqueous layer) surface is smooth and stable with adequate tear volume and that the tear lipid layer is present in adequate thickness.

[Cornea transplant].

PubMed

Garralda, A; Epelde, A; Iturralde, O; Compains, E; Maison, C; Altarriba, M; Goldaracena, M B; Maraví-Poma, E

2006-01-01

The keratoplasty, or cornea transplant, is one of the oldest surgical techniques in opthalmology, whose indication are: 1) tectonic, in order to preserve corneal anatomy and integrity; 2) clinical, in order to eliminate the inflamed corneal tissue in cases refractory to medical treatment; 3) optical, in order to improve visual acuity; and 4) cosmetic, in order to improve the appearance of the eye. Improvements in technique and instruments, as well as in post-operative treatment and the means of preserving donated tissue, have improved survival of the grafts. The Pamplona Model of transplant coordination of the Virgen del Camino Hospital is considered to be original and unique in Spain. The logistics of this program include the protocol for detection and extraction of corneas as well as for keratoplasties.

Applications of biomaterials in corneal wound healing.

PubMed

Tsai, I-Lun; Hsu, Chih-Chien; Hung, Kuo-Hsuan; Chang, Chi-Wen; Cheng, Yung-Hsin

2015-04-01

Disease affecting the cornea is a common cause of blindness worldwide. To date, the amniotic membrane (AM) is the most widely used clinical method for cornea regeneration. However, donor-dependent differences in the AM may result in variable clinical outcomes. To overcome this issue, biomaterials are currently under investigation for corneal regeneration in vitro and in vivo. In this article, we highlight the recent advances in hydrogels, bioengineered prosthetic devices, contact lenses, and drug delivery systems for corneal regeneration. In clinical studies, the therapeutic effects of biomaterials, including fibrin and collagen-based hydrogels and silicone contact lenses, have been demonstrated in damaged cornea. The combination of cells and biomaterials may provide potential treatment in corneal wound healing in the future. Copyright © 2014. Published by Elsevier Taiwan.

The toxic effect of different concentrations of povidone iodine on the rabbit's cornea.

PubMed

Jiang, Jin; Wu, Miaoqin; Shen, Ting

2009-01-01

To investigate the effects of polyvinylpyrrolidone (i.e., povidone) iodine (PVP-I) in different concentrations on the rabbit's cornea. PVP-I in various concentrations was instilled into the conjunctival sac and injected into the anterior chamber independently. The toxicity to the cornea was assessed by fluorescence dye, specular microscopy, and corneal pachymetry. After conjunctival sac PVP-I instillation, severe epithelial damage was observed at the concentrations of 5.0% and 2.5%. After anterior chamber PVP-I injection, significant corneal edema was observed at the concentrations of 2.0% and 1.5%. It can be safe and feasible to use 0.5% or 1.0% concentrations of PVP-I in conjunctival sac instillation for preoperative antisepsis.

Tracing the fate of limbal epithelial progenitor cells in the murine cornea.

PubMed

Di Girolamo, N; Bobba, S; Raviraj, V; Delic, N C; Slapetova, I; Nicovich, P R; Halliday, G M; Wakefield, D; Whan, R; Lyons, J G

2015-01-01

Stem cell (SC) division, deployment, and differentiation are processes that contribute to corneal epithelial renewal. Until now studying the destiny of these cells in a living mammal has not been possible. However, the advent of inducible multicolor genetic tagging and powerful imaging technologies has rendered this achievable in the translucent and readily accessible murine cornea. K14CreER(T2)-Confetti mice that harbor two copies of the Brainbow 2.1 cassette, yielding up to 10 colors from the stochastic recombination of fluorescent proteins, were used to monitor K-14(+) progenitor cell dynamics within the corneal epithelium in live animals. Multicolored columns of cells emerged from the basal limbal epithelium as they expanded and migrated linearly at a rate of 10.8 µm/day toward the central cornea. Moreover, the permanent expression of fluorophores, passed on from progenitor to progeny, assisted in discriminating individual clones as spectrally distinct streaks containing more than 1,000 cells within the illuminated area. The centripetal clonal expansion is suggestive that a single progenitor cell is responsible for maintaining a narrow corridor of corneal epithelial cells. Our data are in agreement with the limbus as the repository for SC as opposed to SC being distributed throughout the central cornea. This is the first report describing stem/progenitor cell fate determination in the murine cornea using multicolor genetic tracing. This model represents a powerful new resource to monitor SC kinetics and fate choice under homeostatic conditions, and may assist in assessing clonal evolution during corneal development, aging, wound-healing, disease, and following transplantation. © 2014 AlphaMed Press.

Occurrence of viral DNA in paired samples of corneal rim and cornea preservation fluid.

PubMed

Broniek, G; Langwińska-Wośko, E; Sybilska, M; Szaflik, J P; Przybylski, M; Wróblewska, M

2017-04-01

Corneal transplants have one of the highest success rates among all transplantological procedures. Corneas intended for transplantation are stored in a preservation fluid, which is then tested for bacterial and fungal infections. Among all analyses of infectious complications following corneal transplants, infections caused by bacteria or fungi are the most prominent. Surprisingly, however, apart from a few publications, there is a lack of data regarding the occurrence of viruses in donor corneas and the risk of transmitting these to their recipients. The intention of this research was therefore to determine the frequency with which human herpesvirus 1 (HHV-1), human herpesvirus 2 (HHV-2), and human adenovirus (HAdV) occur in transplanted corneal tissue, as well as in samples of preservation fluid. The study comprised 57 paired samples, with each pair consisting of a fragment of the corneal tissue remaining after its trepanation for transplantation surgery and a sample of corneal preservation fluid. Sample pairs were all tested for the presence of the DNA of three viruses (HHV-1, HHV-2, and HAdV) using real time PCR technique. Viral DNA was found in three of the tested corneas-HHV-1 DNA in one paired sample (1.8%) and adenovirus DNA in two single samples (3.5%). We postulate that virological testing of corneas for transplantation should be considered, particularly in the case of donors with increased risk factors for herpesvirus and adenovirus reactivation. J. Med. Virol. 89:732-736, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Adaptive differences in the structure and macromolecular compositions of the air and water corneas of the “four-eyed” fish (Anableps anableps)

PubMed Central

SWAMYNATHAN, SHIVALINGAPPA K.; CRAWFORD, MARY A.; ROBISON, W. GERALD; KANUNGO, JYOTSHNABALA; PIATIGORSKY, JORAM

2018-01-01

The water meniscus bisects the eyes of the “four-eyed” fish Anableps anableps, resulting in simultaneous vision in air and water. We compare the structure and macromolecular compositions of the Anableps dorsal (air) and ventral (water) corneas with the fully aquatic zebrafish cornea. The Anableps dorsal corneal epithelium is thicker (>20 cell layers), flatter (~1.94 mm radius of curvature), and contains ~15-fold more glycogen (0.16 μg/μg water-soluble protein) than the ventral corneal epithelium (5–7 cell layers; ~1.63 mm radius of curvature; 0.01 μg glycogen/μg water-soluble protein), which resembles the zebrafish corneal epithelium. Gelsolin is the major water-soluble protein in the zebrafish (~50%) and Anableps dorsal (~38%) and ventral (~21%) corneal epithelia, suggesting that gelsolin was recruited for high corneal expression before these two species diverged at least 100 million years ago and that abundant corneal gelsolin is not limited to aquatic vision. Anableps gelsolin, deduced from its cDNA, is 57% identical to zebrafish gelsolin. Paucity of Anableps corneal F-actin (consistent with high gelsolin) was confirmed by the absence of rhodaminephalloidin staining. We suggest amphibious refraction and protection from UV irradiation and desiccation in air as selective constraints for the specializations of the Anableps dorsal cornea.—Swamynathan, S. K., Craw-ford, M. A., Robison, W. G., Jr., Kanungo, J., Piatigorsky, J. Adaptive differences in the structure and macromolecular compositions of the air and water corneas of the “four-eyed” fish (Anableps anableps). PMID:14597669

Ocular HSV-1: Is the Cornea a Reservoir for Viral Latency or a Fast Pit Stop?

PubMed Central

Kennedy, David P.; Clement, Christian; Arceneaux, Richard L.; Bhattacharjee, Partha S.; Huq, Tashfin S.; Hill, James M.

2010-01-01

Purpose To present a review supporting and refuting evidence from mouse, rabbit, non-human primate, and human studies of herpes simplex virus type 1 (HSV-1) concerning corneal latency. Methods More than 50 research papers on HSV-1 published in peer-reviewed journals were examined. Results Infectious HSV-1 has been found in mouse denervated tissues and in tissues with negative cultures from the corresponding ganglion. However, the different mouse strains have shown varied responses to different strains of HSV, making it difficult to relate such findings to humans. Rabbit studies provide excellent evidence for HSV-1 corneal latency including data on HSV-1 migration from the cornea into the corneoscleral rim and on the distribution of HSV-1 DNA in the cornea. However, the available methods for the detection of infectious HSV-1 may not be sensitive enough to detect low-level infection. Infectious HSV-1 has been successfully isolated from the tears of non-human primates in the absence of detectable corneal lesions. The recurrence of corneal ulcers in non-human primates before the appearance of infectious HSV-1 in tears suggests that the origin of the HSV-1 is the cornea, rather than the TG. Human studies presented evidence of both ganglion and corneal latency. Conclusion Understanding HSV-1 disease progression and the possibility of corneal latency could lead to more effective treatments for herpetic keratitis. However, it is unlikely that operational latency in the cornea will be definitively proven unless a new method with higher sensitivity for the detection of infectious virus is developed. PMID:21304287

Ultrastructure and mineral composition of the cornea cuticle in the compound eyes of a supralittoral and a marine isopod.

PubMed

Alagboso, Francisca I; Reisecker, Christian; Hild, Sabine; Ziegler, Andreas

2014-08-01

The cuticle of the cornea in Crustacea is an interesting example of a composite material compromising between two distinct functions. As part of the dioptric apparatus of the ommatidia within the complex eye it forms transparent micro-lenses that should as well maintain the mechanical stability of the head capsule. We analyzed the ultrastructure and composition of the isopod cornea cuticle of the terrestrial species Ligia oceanica and the marine Sphaeroma serratum. We used a variety of tissue preparation methods, electron microscopic techniques as well as electron microprobe analysis and Raman spectroscopic imaging. The results reveal various structural adaptations that likely increase light transmission. These are an increase in the thickness of the epicuticle, a reduction of the thickness of the outer layer of calcite, a spatial restriction of pore canals to interommatidial regions, and, for S. serratum only, an increase in calcite crystal size. In both species protein-chitin fibrils within the proximal exocuticle form a peculiar reticular structure that does not occur within the cuticle of the head capsule. In L. oceanica differential mineralization results in a spherically shaped interface between mineralized and unmineralized endocuticle, likely an adaptation to increase the refractive power of the cornea maintaining the mechanical stability of the cuticle between the ommatidia. The results show that the habitat and differences in the general structure of the animal's cuticle affect the way in which the cornea is adapted to its optical function. Copyright © 2014 Elsevier Inc. All rights reserved.

Cornea and anterior eye assessment with placido-disc keratoscopy, slit scanning evaluation topography and scheimpflug imaging tomography

PubMed Central

Martin, Raul

2018-01-01

Current corneal assessment technologies make the process of corneal evaluation extremely fast and simple and several devices and technologies allow to explore and to manage patients. The purpose of this special issue is to present and also to update in the evaluation of cornea and ocular surface and this second part, reviews a description of the corneal topography and tomography techniques, providing updated information of the clinical recommendations of these techniques in eye care practice. Placido-based topographers started an exciting anterior corneal surface analysis that allows the development of current corneal tomographers that provide a full three-dimensional reconstruction of the cornea including elevation, curvature, and pachymetry data of anterior and posterior corneal surfaces. Although, there is not an accepted reference standard technology for corneal topography description and it is not possible to determine which device produces the most accurate topographic measurements, placido-based topographers are a valuable technology to be used in primary eye care and corneal tomograhers expanding the possibilities to explore cornea and anterior eye facilitating diagnosis and follow-up in several situations, raising patient follow-up, and improving the knowledge regarding to the corneal anatomy. Main disadvantages of placido-based topographers include the absence of information about the posterior corneal surface and limited corneal surface coverage without data from the para-central and/or peripheral corneal surface. However, corneal tomographers show repeatable anterior and posterior corneal surfaces measurements, providing full corneal thickness data improving cornea, and anterior surface assessment. However, differences between devices suggest that they are not interchangeable in clinical practice. PMID:29480244

The corneal transplant score: a simple corneal graft candidate calculator.

PubMed

Rosenfeld, Eldar; Varssano, David

2013-07-01

Shortage of corneas for transplantation has created long waiting lists in most countries. Transplant calculators are available for many organs. The purpose of this study is to describe a simple automatic scoring system for keratoplasty recipient candidates, based on several parameters that we consider most relevant for tissue allocation, and to compare the system's accuracy in predicting decisions made by a cornea specialist. Twenty pairs of candidate data were randomly created on an electronic spreadsheet. A single priority score was computed from the data of each candidate. A cornea surgeon and the automated system then decided independently which candidate in each pair should have surgery if only a single cornea was available. The scoring system can calculate values between 0 (lowest priority) and 18 (highest priority) for each candidate. Average score value in our randomly created cohort was 6.35 ± 2.38 (mean ± SD), range 1.28 to 10.76. Average score difference between the candidates in each pair was 3.12 ± 2.10, range 0.08 to 8.45. The manual scoring process, although theoretical, was mentally and emotionally demanding for the surgeon. Agreement was achieved between the human decision and the calculated value in 19 of 20 pairs. Disagreement was reached in the pair with the lowest score difference (0.08). With worldwide donor cornea shortage, waiting for transplantation can be long. Manual sorting of priority for transplantation in a long waiting list is difficult, time-consuming and prone to error. The suggested system may help achieve a justified distribution of available tissue.

Cornea and anterior eye assessment with placido-disc keratoscopy, slit scanning evaluation topography and scheimpflug imaging tomography.

PubMed

Martin, Raul

2018-03-01

Current corneal assessment technologies make the process of corneal evaluation extremely fast and simple and several devices and technologies allow to explore and to manage patients. The purpose of this special issue is to present and also to update in the evaluation of cornea and ocular surface and this second part, reviews a description of the corneal topography and tomography techniques, providing updated information of the clinical recommendations of these techniques in eye care practice. Placido-based topographers started an exciting anterior corneal surface analysis that allows the development of current corneal tomographers that provide a full three-dimensional reconstruction of the cornea including elevation, curvature, and pachymetry data of anterior and posterior corneal surfaces. Although, there is not an accepted reference standard technology for corneal topography description and it is not possible to determine which device produces the most accurate topographic measurements, placido-based topographers are a valuable technology to be used in primary eye care and corneal tomograhers expanding the possibilities to explore cornea and anterior eye facilitating diagnosis and follow-up in several situations, raising patient follow-up, and improving the knowledge regarding to the corneal anatomy. Main disadvantages of placido-based topographers include the absence of information about the posterior corneal surface and limited corneal surface coverage without data from the para-central and/or peripheral corneal surface. However, corneal tomographers show repeatable anterior and posterior corneal surfaces measurements, providing full corneal thickness data improving cornea, and anterior surface assessment. However, differences between devices suggest that they are not interchangeable in clinical practice.

Three-dimensional modeling of metabolic species transport in the cornea with a hydrogel intrastromal inlay.

PubMed

Pinsky, Peter M

2014-05-15

Intrastromal inlays for refractive correction of presbyopia are being adopted into clinical practice. An important concern is the effect of the inlay on the long-term health of the cornea due to disturbances in the concentration profiles of metabolic species. A three-dimensional metabolic model for the cornea is employed to investigate oxygen, glucose, and lactate ion transport in the cornea and to estimate changes in species concentrations induced by the introduction of a hydrogel inlay. A reaction-diffusion metabolic model, appropriate for highly oxygen-permeable hydrogel inlays, is used to describe cellular consumption of oxygen and glucose and production of lactic acid. A three-layer corneal geometry (epithelium, stroma, endothelium) is employed with a hydrogel inlay placed under a lamellar flap. The model is solved numerically by the finite element method. For a commercially available hydrogel material with a relative inlay diffusivity of 43.5%, maximum glucose depletion and lactate ion accumulation occur anterior to the inlay and both are less than 3%. Below 20% relative diffusivity, glucose depletion and lactate ion accumulation increase exponentially. Glucose depletion increases slightly with increasing depth of inlay placement. The flux of metabolic species is modified by an inlay, depending on the inlay relative diffusivity. For commercially available hydrogel materials and a typical inlay design, predicted changes in species concentrations are small when compared to the variation of concentrations across the normal cornea. In general, glucose depletion and lactate ion accumulation are highly sensitive to inlay diffusivity and somewhat insensitive to inlay depth. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

Transmission Electron Microscopy Analysis of Epithelial Basement Membrane Repair in Rabbit Corneas With Haze

PubMed Central

Torricelli, Andre A. M.; Singh, Vivek; Agrawal, Vandana; Santhiago, Marcony R.; Wilson, Steven E.

2013-01-01

Purpose. To assess the ultrastructure of the epithelial basement membrane using transmission electron microscopy (TEM) in rabbit corneas with and without subepithelial stroma opacity (haze). Methods. Two groups of eight rabbits each were included in this study. Photorefractive keratectomy (PRK) was performed using an excimer laser. The first group had −4.5-diopter (−4.5D) PRK and the second group had −9.0D PRK. Contralateral eyes were unwounded controls. Rabbits were sacrificed at 4 weeks after surgery. Immunohistochemical analysis was performed to detect the myofibroblast marker α-smooth muscle actin (SMA). TEM was performed to analyze the ultrastructure of the epithelial basement membrane and stroma. Results. At 4 weeks after PRK, α-SMA+ myofibroblasts were present at high density in the subepithelial stroma of rabbit eyes that had −9.0D PRK, along with prominent disorganized extracellular matrix, whereas few myofibroblasts and little disorganized extracellular matrix were noted in eyes that had −4.5D PRK. The epithelial basement membrane was irregular and discontinuous and lacking typical morphology in all corneas at 1 month after −9D PRK compared to corneas at 1 month in the −4.5D PRK group. Conclusions. The epithelial basement membrane acts as a critical modulator of corneal wound healing. Structural and functional defects in the epithelial basement membrane correlate to both stromal myofibroblast development from precursor cells and continued myofibroblast viability, likely through the modulation of epithelial–stromal interactions mediated by cytokines. Prolonged stromal haze in the cornea is associated with abnormal regeneration of the epithelial basement membrane. PMID:23696606

Finite element analysis of blunt foreign body impact on the cornea after PRK and LASIK.

PubMed

Mousavi, Seyed Jamaleddin; Nassiri, Nariman; Masoumi, Nafiseh; Nassiri, Nader; Majdi-N, Mercede; Farzaneh, Solmaz; Djalilian, Ali R; Peyman, Gholam A

2012-01-01

To investigate the effect of blunt foreign body impact on a human cornea after photorefractive keratectomy (PRK) and LASIK using a simulation model. Computational simulations were performed using a finite element analysis program (LS-Dyna, Livermore Software Technology Corp). The blunt foreign body was set to impact at the center of the corneal surface models (after PRK and LASIK) with thicknesses of 500, 450, 400, 350, and 300 μm. Corneal rupture was assumed to occur at a peak stress of 9.45 MPa and at a strain of 18%. The foreign body projectile was blunt in shape, made from aluminum, contained plastic-kinematic properties, and had a density of 2700 kg/m(3). The projectile was launched at the center of the cornea with velocities ranging from 20 to 60 m/s. The threshold of impact velocities creating rupture in corneal thicknesses of 500, 450, 400, 350, and 300 μm were 33, 32.8, 30.7, 27.9, and 22.8 m/s, respectively, in the PRK model. In the LASIK model, the thresholds creating rupture in the stromal bed of the corneas with thicknesses of 500, 450, 400, 350, and 300 μm were 40, 38.1, 35.6, 31.5, and 26.7 m/s, respectively. The 110-μm corneal flap in the LASIK model ruptured at all velocities. Ruptures occurred at lower velocities in the PRK cornea model than in the corneal stromal bed of the LASIK model following blunt foreign body impact. Copyright 2012, SLACK Incorporated.

Cold chain monitoring during cold transportation of human corneas for transplantation.

PubMed

Net, M; Trias, E; Navarro, A; Ruiz, A; Diaz, P; Fontenla, J R; Manyalich, M

2003-08-01

As recommended by international standards the cornea should be maintained in a specific temperature range (2 degrees -8 degrees C) to guarantee its viability. However, there is no standard packaging method to maintain these conditions during transport. Our packaging system is similar to those used by the main eye banks in Spain and elsewhere in Europe. The objective is to monitor the cold chain in the current packaging method to validate the maintenance of temperature within the adequate range for a minimum 24-hour period. The effects of the following variables were studied: number and freezing temperature of the cold packs; air volume in the packaging system; position of the cornea in the packaging system; and the wall section of the container. Exterior temperature was maintained constant at 20 degrees to 24 degrees C. The cold chain was monitored using a device that measures temperature continuously and for which a histogram of temperature variation can be downloaded to a computer for further analysis. When the cold packs were frozen to -40 degrees C or the number of cold packs increased to four, the temperature decreased quickly to 0 degrees C and the transport period was not prolonged. The main objective was to improve isolation by reducing inner air volume, and maintaining the position of the cornea in the container. The currently used cold packaging systems (not frozen, 4 degrees C) do not maintain the temperature within the accepted range for the required distribution period. The improved system maintains the cornea at between 2 degrees C and 6 degrees C for a minimum of 24 hours.

Optically inspired biomechanical model of the human eyeball.

PubMed

Sródka, Wieslaw; Iskander, D Robert

2008-01-01

Currently available biomechanical models of the human eyeball focus mainly on the geometries and material properties of its components while little attention has been given to its optics--the eye's primary function. We postulate that in the evolution process, the mechanical structure of the eyeball has been influenced by its optical functions. We develop a numerical finite element analysis-based model in which the eyeball geometry and its material properties are linked to the optical functions of the eye. This is achieved by controlling in the model all essential optical functions while still choosing material properties from a range of clinically available data. In particular, it is assumed that in a certain range of intraocular pressures, the eye is able to maintain focus. This so-called property of optical self-adjustments provides a more constrained set of numerical solutions in which the number of free model parameters significantly decreases, leading to models that are more robust. Further, we investigate two specific cases of a model that satisfies optical self-adjustment: (1) a full model in which the cornea is flexibly attached to sclera at the limbus, and (2) a fixed cornea model in which the cornea is not allowed to move at the limbus. We conclude that for a biomechanical model of the eyeball to mimic the optical function of a real eye, it is crucial that the cornea is allowed to move at the limbal junction, that the materials used for the cornea and sclera are strongly nonlinear, and that their moduli of elasticity remain in a very close relationship.

Long-term consequences of topical dexamethasone treatment during acute corneal HSV-1 infection on the immune system

PubMed Central

Chucair-Elliott, Ana J.; Carr, Meghan M.; Carr, Daniel J. J.

2017-01-01

Herpes simplex virus type 1 (HSV-1) is a leading cause of neurotrophic keratitis (NTK). NTK is characterized by decreased corneal sensation from damage to the corneal sensory fibers. We have reported on the regression of corneal nerves and their function during acute HSV-1 infection. That nerve loss is followed by an aberrant process of nerve regeneration during the latent phase of infection that lacks functional recovery. We recently showed the elicited immune response in the infected cornea, and not viral replication itself, is part of the mechanism responsible for the nerve degeneration process after infection. Specifically, we showed infected corneas topically treated with dexamethasone (DEX) significantly retained both structure and sensitivity of the corneal nerve network in comparison to mice treated with control eye drops, consistent with decreased levels of proinflammatory cytokines and reduced influx of macrophages and CD8+ T cells into the cornea. This study was undertaken to analyze the long-term effect of such a localized, immunosuppressive paradigm (DEX drops on the cornea surface during the first 8 d of HSV-1 infection) on the immune system and on corneal pathology. We found the profound immunosuppressive effect of DEX on lymphoid tissue was sustained in surviving mice for up to 30 d postinfection (p.i.). DEX treatment had prolonged effects, preserving corneal innervation and its function and blunting neovascularization, as analyzed at 30 d p.i. Our data support previously reported observations of an association between the persistent presence of inflammatory components in the latently infected cornea and structural and functional nerve defects in NTK. PMID:28115476

Assessing the Impact of Diabetes Mellitus on Donor Corneal Endothelial Cell Density.

PubMed

Liaboe, Chase A; Aldrich, Benjamin T; Carter, Pamela C; Skeie, Jessica M; Burckart, Kimberlee A; Schmidt, Gregory A; Reed, Cynthia R; Zimmerman, M Bridget; Greiner, Mark A

2017-05-01

To quantify changes in endothelial cell density (ECD) of donor corneal tissue in relation to the presence or absence of a medical history of diabetes mellitus diagnosis, treatment, and complications. A retrospective review was performed for all corneas collected at Iowa Lions Eye Bank between January 2012 and December 2015. For purposes of analysis, donor corneas were divided into 4 groups: nondiabetic, non-insulin-dependent diabetic, insulin-dependent diabetic without medical complications due to diabetes, and insulin-dependent diabetic with medical complications due to diabetes. ECD values (obtained through specular microscopy) and transplant suitability for endothelial transplantation (determined by the standard protocol of the eye bank) were compared among groups using linear mixed model analysis. In total, 4185 corneas from 2112 donors were included for analysis. Insulin-dependent diabetic samples with medical complications due to diabetes (N = 231 from 119 donors) showed lower ECD values compared with nondiabetic samples (-102 cells/mm, P = 0.049) and non-insulin-dependent diabetic samples (-117 cells/mm, P = 0.031). ECD values did not differ significantly among the remaining groups. The likelihood of suitability for endothelial transplantation did not differ among all 4 groups. Corneas from donors with insulin-dependent diabetes mellitus and medical complications resulting from the disease have lower mean ECD values compared with other donors. However, our analysis suggests that these corneas are equally likely to be included in the donor pool for corneal transplantation. Additional studies are needed to determine the mechanism(s) contributing to cell loss in donors with advanced diabetes and to assess associated endothelial cell functional impairment.

In vivo confocal Raman spectroscopy of the human cornea.

PubMed

Bauer, N J; Hendrikse, F; March, W F

1999-07-01

To investigate the feasibility of a confocal Raman spectroscopic technique for the noninvasive assessment of corneal hydration in vivo in two legally blind subjects. A laser beam (632.8 nm; 15 mJ) was maintained on the cornea by using a microscope objective lens (x25 magnification, NA = 0.5, f = 10 mm) both for focusing the incident light as well as collecting the Raman backscattered light, in a 180 degrees backscatter configuration. An optical fiber, acting as the confocal pinhole for elimination of light from out-of-focus places, was coupled to a spectrometer that dispersed the collected light onto a sensitive array detector for rapid spectral data acquisition over a range from 2,890 to 3,590/cm(-1). Raman spectra were recorded from the anterior 100-150 microm of the cornea over a period before and after topical application of a mild dehydrating solution. The ratio between the amplitudes of the signals at 3,400/cm(-1) (OH-vibrational mode of water) and 2,940/cm(-1) (CH-vibrational mode of proteins) was used as a measure for corneal hydration. High signal-to-noise ratio (SNR = 25) Raman spectra were obtained from the human corneas by using 15 mJ of laser light energy. Qualitative changes in the hydration of the anteriormost part of the corneas could be observed as a result of the dehydrating agent. With adequate improvements in system safety, confocal Raman spectroscopy could potentially be applied clinically as a noninvasive tool for the assessment of corneal hydration in vivo.

Epifluorescence Intravital Microscopy of Murine Corneal Dendritic Cells

PubMed Central

Rosenbaum, James T.; Planck, Stephen R.

2010-01-01

Purpose. Dendritic cells (DCs) are antigen-presenting cells vital for initiating immune responses. In this study the authors examined the in vivo migratory capability of resident corneal DCs to various stimuli. Methods. The authors used mice expressing enhanced yellow fluorescent protein (eYFP) under control of the CD11c promoter to visualize corneal DCs. To assess the distribution and mobility of DCs, normal corneas were imaged in vivo and ex vivo with fluorescence microscopy. Intravital microscopy was used to examine the responses of resident central and peripheral corneal DCs to silver nitrate injury, lipopolysaccharide, microspheres, and tumor necrosis factor (TNF-α). In some experiments, TNF-α injection was used to first induce centripetal migration of DCs to the central cornea, which was subsequently reinjected with microspheres. Results. In normal corneas, DCs were sparsely distributed centrally and were denser in the periphery, with epithelial-level DCs extending into the epithelium. Videomicroscopy showed that though cell processes were in continuous movement, cells generally did not migrate. Within the first 6 hours after stimulation, neither central nor peripheral corneal DCs exhibited significant lateral migration, but central corneal DCs assumed extreme morphologic changes. An increased number of DCs in the TNF-α–stimulated central cornea were responsive to subsequent microsphere injection by adopting a migratory behavior, but not with increased speed. Conclusions. In vivo imaging reveals minimal lateral migration of corneal DCs after various stimuli. In contrast, DCs within the central cornea after initial TNF-α injection are more likely to respond to a secondary insult with lateral migration. PMID:20007837

Eye development and the appearance and maintenance of corneal transparency

NASA Technical Reports Server (NTRS)

Conrad, G. W.; Funderburgh, J. L.; Spooner, B. S. (Principal Investigator)

1992-01-01

Embryonic development of the eye, including the cornea, depends on the appearance and steady maintenance of intraocular pressure. The eye is a gravity-sensitive organ, as evidence by changes in pupil diameter during parabolic flight. The cornea is largely a paracrystal of extracellular matrix. The extent to which it will polymerize normally in microgravity has yet to be determined.

[Current approaches to evaluating the anatomic and functional status of the cornea].

PubMed

Avetisov, S E; Borodina, N V; Kobzova, M V; Musaeva, G M

2010-01-01

The review provides data on current methods for evaluating the anatomic and functional status of the cornea (light refraction, light transmission, and biomechanical properties, in particular). It analyzes the main advantages and disadvantages of basic (biomicroscopy, endothelial microscopy, ophthalmometry, topography, and pachymetry) and special (confocal microscopy, optical coherence tomography, ultrasound biomicroscopy, aberrometry, bidirectional corneal applanation, and keratoesthesiometry) studies.

Diffusion and Monod kinetics model to determine in vivo human corneal oxygen-consumption rate during soft contact lens wear.

PubMed

Del Castillo, Luis F; da Silva, Ana R Ferreira; Hernández, Saul I; Aguilella, M; Andrio, Andreu; Mollá, Sergio; Compañ, Vicente

2015-01-01

We present an analysis of the corneal oxygen consumption Qc from non-linear models, using data of oxygen partial pressure or tension (P(O2) ) obtained from in vivo estimation previously reported by other authors. (1) METHODS: Assuming that the cornea is a single homogeneous layer, the oxygen permeability through the cornea will be the same regardless of the type of lens that is available on it. The obtention of the real value of the maximum oxygen consumption rate Qc,max is very important because this parameter is directly related with the gradient pressure profile into the cornea and moreover, the real corneal oxygen consumption is influenced by both anterior and posterior oxygen fluxes. Our calculations give different values for the maximum oxygen consumption rate Qc,max, when different oxygen pressure values (high and low P(O2)) are considered at the interface cornea-tears film. Present results are relevant for the calculation on the partial pressure of oxygen, available at different depths into the corneal tissue behind contact lenses of different oxygen transmissibility. Copyright © 2014. Published by Elsevier Espana.

Latest result of PRK with excimer laser

NASA Astrophysics Data System (ADS)

Okamoto, Shinseiro; Okamoto, Michika

1996-05-01

We have in the last two years, performed PRK operation on over 300 human myopic eyes using ArF excimer laser with a Summit 'Omnimed' machine. For the initial 53 myopic eyes we treated, results were very good for those with correction less than minus 6 diopters. However, as previously reported, we also witnessed some regression for those eyes exceeding correction of more than minus 6 diopters. To counter such ill results of PRK we devised and suggested many new procedures for PRK with very good results. One such invention is the 'Okamoto-type' cooling machine for the cornea which reduces and stabilizes cornea temperature at 0 degrees Celsius while simultaneously bathing the cornea with special cooling fluid. After the operation, EGF, fibronectin and hexapeptide were administered using eyedrops. Soft contact lenses were used to protect the cornea, improve delivery of medication to the operated area, prevent infection and inflammation and also promote uniform and faster ephiterium regrowth. We were able to document very good post-operative results using this method, thereby giving us strong assurance that we have reached a significant milestone in PRK operation. Our report today covers post operative results of the 52 eyes we operated on and tracked for more than one year.

Transient viscous response of the human cornea probed with the Surface Force Apparatus.

PubMed

Zappone, Bruno; Patil, Navinkumar J; Lombardo, Marco; Lombardo, Giuseppe

2018-01-01

Knowledge of the biomechanical properties of the human cornea is crucial for understanding the development of corneal diseases and impact of surgical treatments (e.g., corneal laser surgery, corneal cross-linking). Using a Surface Force Apparatus we investigated the transient viscous response of the anterior cornea from donor human eyes compressed between macroscopic crossed cylinders. Corneal biomechanics was analyzed using linear viscoelastic theory and interpreted in the framework of a biphasic model of soft hydrated porous tissues, including a significant contribution from the pressurization and viscous flow of fluid within the corneal tissue. Time-resolved measurements of tissue deformation and careful determination of the relaxation time provided an elastic modulus in the range between 0.17 and 1.43 MPa, and fluid permeability of the order of 10-13 m4/(N∙s). The permeability decreased as the deformation was increased above a strain level of about 10%, indicating that the interstitial space between fibrils of the corneal stromal matrix was reduced under the effect of strong compression. This effect may play a major role in determining the observed rate-dependent non-linear stress-strain response of the anterior cornea, which underlies the shape and optical properties of the tissue.

A boussinesq model of natural convection in the human eye and the formation of Krukenberg's spindle.

PubMed

Heys, Jeffrey J; Barocas, Victor H

2002-03-01

The cornea of the human eye is cooled by the surrounding air and by evaporation of the tear film. The temperature difference between the cornea and the iris (at core body temperature) causes circulation of the aqueous humor in the anterior chamber of the eye. Others have suggested that the circulation pattern governs the shape of the Krukenberg spindle, a distinctive vertical band of pigment on the posterior cornea surface in some pathologies. We modeled aqueous humor flow the human eye, treating the humor as a Boussinesq fluid and setting the corneal temperature based on infrared surface temperature measurements. The model predicts convection currents in the anterior chamber with velocities comparable to those resulting from forced flow through the gap between the iris and lens. When paths of pigment particles are calculated based on the predicted flow field, the particles circulate throughout the anterior chamber but tend to be near the vertical centerline of the eye for a greatest period of time. Further, the particles are usually in close proximity to the cornea only when they are near the vertical centerline. We conclude that the convective flow pattern of aqueous humor is consistent with a vertical pigment spindle.

[The influence of sodium hyaluronate solution and artificial tears on higher-order aberrations].

PubMed

Yamashita, Tsutomu; Ochi, Shintarou; Inoue, Yasushi; Miki, Atsushi; Kiryu, Junichi; Tabuchi, Akio

2013-12-01

To investigate the influence of sodium hyaluronate solution (HA) and artificial tears (AT) on higher-order aberrations (HOAs). Twenty four eyes of 24 normal subjects and 11 eyes of 11 dry eye patients were examined. Cornea and ocular wavefront aberrations (total, spherical-like and coma-like) were measured with a Hartmann-Shack wavefront aberrometer before and after 0.1% or 0.3% HA, AT. The consecutively obtained data of the cornea and ocular HOAs were analyzed in the central 4-mm diameter for coma-like, spherical-like and total HOAs. Average HOAs, as well as fluctuation index (FI) and stability index (SI) of the HOAs over time were compared between the two groups. In normal subjects, the AVE of all aberration parameters and FI showed an increase depending on viscosity of the HA (p < 0.001). After AT and 0.1% HA treatment the cornea aberration of the dry eye patients changed from a sawtooth pattern to a stable pattern. Cornea HOAs decreased, and the optical characteristics showed improvement after AT and 0.1% HA in the dry eye patients. HOAs increased depending on the viscosity of the HA, and optical stability worsened.

In-vivo immunofluorescence confocal microscopy of herpes simplex virus type 1 keratitis

NASA Astrophysics Data System (ADS)

Kaufman, Stephen C.; Laird, Jeffery A.; Beuerman, Roger W.

1996-05-01

The white-light confocal microscope offers an in vivo, cellular-level resolution view of the cornea. This instrument has proven to be a valuable research and diagnostic tool for the study of infectious keratitis. In this study, we investigate the direct visualization of herpes simplex virus type 1 (HSV-1)-infected corneal epithelium, with in vivo confocal microscopy, using HSV-1 immunofluorescent antibodies. New Zealand white rabbits were infected with McKrae strain of HSV-1 in one eye; the other eye of each rabbit was used as an uninfected control. Four days later, the rabbits were anesthetized and a cellulose sponge was applied to each cornea, and a drop of direct HSV fluorescein-tagged antibody was placed on each sponge every 3 to 5 minutes for 1 hour. Fluorescence confocal microscopy was then performed. The HSV-infected corneas showed broad regions of hyperfluorescent epithelial cells. The uninfected corneas revealed no background fluorescence. Thus, using the confocal microscope with a fluorescent cube, we were able to visualize HSV-infected corneal epithelial cells tagged with a direct fluorescent antibody. This process may prove to be a useful clinical tool for the in vivo diagnosis of HSV keratitis.

A DNA fragment of Leptospira interrogans encodes a protein which shares epitopes with equine cornea.

PubMed

Lucchesi, P M; Parma, A E

1999-11-30

Horses infected with Leptospira interrogans present several clinical disorders, one of them being recurrent uveitis. An antigenic relationship between this bacterium and equine cornea has been described in previous studies. With the aim to make progress on defining the molecular basis and pathogenesis of equine recurrent uveitis, here we describe the cloning of one DNA fragment from a Leptospira interrogans serovar pomona genomic lambda gt11 library. Although there are references of transcription of leptospiral genes in E. coli from their own leptospiral promoters, in this recombinant construction the leptospiral DNA was located under the control of lacZ promoter since no expression could be detected in the absence of IPTG. This clone, isolated by expression screening with polyclonal serum raised against equine corneal proteins, encodes a 90 kDa protein of L. interrogans which crossreacts with equine cornea as proved Western-blotting. Antibodies directed against this leptospiral protein strongly recognised a 66 kDa equine corneal protein, one of those recognised by an anti-equine cornea serum. Our findings suggest that an immune response to 90 kDa protein participates in pathogenesis of equine uveitis.

Three-dimensional and topographic relationships between the orbital margins with reference to assessment of eyeball protrusion.

PubMed

Shin, Kang-Jae; Lee, Shin-Hyo; Koh, Ki-Seok; Song, Wu-Chul

2017-03-01

This study investigated the topographic relationships among the eyeball and four orbital margins with the aim of identifying the correlation between orbital geometry and eyeball protrusion in Koreans. Three-dimensional (3D) volume rendering of the face was performed using serial computed-tomography images of 141 Koreans, and several landmarks on the bony orbit and the cornea were directly marked on the 3D volumes. The anterior-posterior distances from the apex of the cornea to each orbital margin and between the orbital margins were measured in both eyes. The distances from the apex of the cornea to the superior, medial, inferior, and lateral orbital margins were 5.8, 5.8, 12.0, and 17.9 mm, respectively. Differences between sides were observed in all of the orbital margins, and the distances from the apex of the cornea to the superior and inferior orbital margins were significantly greater in females than in males. The anterior-posterior distance between the superior and inferior orbital margins did not differ significantly between males (6.3 mm) and females (6.2 mm). The data obtained in this study will be useful when developing practical guidelines applicable to forensic facial reconstruction and orbitofacial surgeries.

Investigation of the morphology of the features generated via femtosecond lasers in the interior of a bovine cornea sections

NASA Astrophysics Data System (ADS)

Vukelic, Sinisa; Kongsuwan, Panjawat; Yao, Y. Lawrence

2013-02-01

Nonlinear absorption of femtosecond laser pulses enables the induction of bubble cavities in the interior of eye cornea without affecting other parts of an eye, a phenomena utilized for flap formation in laser assisted corneal surgery. In the present study laser pulses were focused in the interior of the sections of bovine cornea. Tight focus of the laser pulses results in the plasma formation followed by its explosive expansion, which drives cavity formation. The morphology of the generated features as well as the nature of the physical mechanisms of the phenomenon as a function of process parameters is discussed. Numerical model is proposed to develop predictive capabilities for the feature size and shape and the results are compared against the experimental findings.

Epidermal growth factor in alkali-burned corneal epithelial wound healing.

PubMed

Singh, G; Foster, C S

1987-06-15

We conducted a double-masked study to evaluate the effect of epidermal growth factor on epithelial wound healing and recurrent erosions in alkali-burned rabbit corneas. Epithelial wounds 10 mm in diameter healed completely under the influence of topical epidermal growth factor, whereas the control corneas did not resurface in the center. On reversal of treatment, the previously nonhealing epithelial defects healed when treated with topical epidermal growth factor eyedrops. Conversely, the epidermal growth factor-treated and resurfaced corneas developed epithelial defects when treatment was discontinued. Histopathologic examination disclosed hyperplastic epithelium growing over the damaged stroma laden with polymorphonuclear leukocytes when treated with epidermal growth factor eyedrops, but it did not adhere to the underlying tissue. Hydropic changes were seen intracellularly as well as between the epithelial cells and the stroma.

Quantitative assessment of corneal viscoelasticity using optical coherence elastography and a modified Rayleigh-Lamb equation

NASA Astrophysics Data System (ADS)

Han, Zhaolong; Aglyamov, Salavat R.; Li, Jiasong; Singh, Manmohan; Wang, Shang; Vantipalli, Srilatha; Wu, Chen; Liu, Chih-hao; Twa, Michael D.; Larin, Kirill V.

2015-02-01

We demonstrate the use of a modified Rayleigh-Lamb frequency equation in conjunction with noncontact optical coherence elastography to quantify the viscoelastic properties of the cornea. Phase velocities of air-pulse-induced elastic waves were extracted by spectral analysis and used for calculating the Young's moduli of the samples using the Rayleigh-Lamb frequency equation (RLFE). Validation experiments were performed on 2% agar phantoms (n=3) and then applied to porcine corneas (n=3) in situ. The Young's moduli of the porcine corneas were estimated to be ˜60 kPa with a shear viscosity ˜0.33 Pa.s. The results demonstrate that the RLFE is a promising method for noninvasive quantification of the corneal biomechanical properties and may potentially be useful for clinical ophthalmological applications.

Application Of The Excimer Laser To Area Recontouring Of The Cornea

NASA Astrophysics Data System (ADS)

Yoder, Paul R.; Telfair, William B.; Warner, John W.; Martin, Clifford A.; L'Esperance, Francis A.

1989-04-01

Excimer lasers operating at 193 nm are being used experimentally in a special type of materials processing wherein the central portion of the anterior surface of the human cornea is selectively ablated so as to change its refractive power and, hopefully, improve impaired vision. Research to date has demonstrated recontouring as a potential means for reducing myopia and hyperopia of cadaver eyes while studies of ablations on the corneas of living monkeys and of blind human volunteers show promise of prompt and successful healing. The procedure has also shown merit in removing superficial scars from the corneal surface. In this paper, we describe the electro-optical system used to deliver the UV laser beam in these experiments and report some preliminary results of the ablation studies.

Imaging of acoustic waves induced by excimer laser ablation of the cornea

NASA Astrophysics Data System (ADS)

Rossi, Francesca; Pini, Roberto; Siano, Salvatore; Salimbeni, Renzo

1996-12-01

In this present study a pump-and-probe imaging set up was arranged to image and analyze the evolution of pressure waves induced by ArF ablation of the cornea, during their propagation into the eyeball. In vitro experiments simulating the effects of clinical PRK have been performed by using an artificial model of the human eyeball, composed of a cell filled with hyaluronic acid gel with a sample of freshly excised bovine cornea placed on the gel surface. LAser irradiation was provided at a fluence of 180 mJ/cm2. Irradiation spot diameters were varied in the range 2.0-5.0 mm. Images of the traveling acoustic waves evidenced diffraction effects, related to the diameter of laser spots on the corneal surface.

Chronic Lunar Dust Exposure on Rat Cornea: Evaluation by Gene Expression Profiling

NASA Technical Reports Server (NTRS)

Theriot, C. A.; Glass, A.; Lam, C-W.; James, J.; Zanello, S. B.

2014-01-01

Lunar dust is capable of entering habitats and vehicle compartments by sticking to spacesuits or other objects that are transferred into the spacecraft from the lunar surface and has been reported to cause irritation upon exposure. During the Apollo missions, crewmembers reported irritation specifically to the skin and eyes after contamination of the lunar and service modules. It has since been hypothesized that ocular irritation and abrasion might occur as a result of such exposure, impairing crew vision. Recent work has shown that both ultrafine and unground lunar dust exhibited minimal irritancy of the ocular surface (i.e., cornea); however, the assessment of the severity of ocular damage resulting from contact of lunar dust particles to the cornea has focused only on macroscopic signs of mechanical irritancy and cytotoxicity. Given the chemical reactive properties of lunar dust, exposure of the cornea may contribute to detrimental effects at the molecular level including but not limited to oxidative damage. Additionally, low level chronic exposures may confound any results obtained in previous acute studies. We report here preliminary results from a tissue sharing effort using 10-week-old Fischer 344 male rats chronically exposed to filtered air or jet milled lunar dust collected during Apollo 14 using a Jaeger-NYU nose-only chamber for a total of 120 hours (6 hours daily, 5 days a week) over a 4-week period. RNA was isolated from corneas collected from rats at 1 day and 7 days after being exposed to concentrations of 0, 20, and 60 mg/m3 of lunar dust. Microarray analysis was performed using the Affymetrix GeneChip Rat Genome 230 2.0 Array with Affymetrix Expression Console and Transcriptome Analysis Console used for normalization and secondary analysis. An Ingenuity iReport"TM" was then generated for canonical pathway identification. The number of differentially expressed genes identified increases with dose compared to controls suggesting a more severe response to the lunar dust insult at higher levels. Pathways of interests that have been identified in all exposed samples include oxidative stress response, mitochondrial dysfunction, fibrosis, epithelial healing, TGF-Beta? signaling, and extracellular matrix remodeling. Several biological processes related to cell migration, cellular proliferation, and eye development were also identified to be altered by exposure to lunar dust. Our preliminary results suggest that even a chronic insult of lunar dust as low as 20 mg/m(exp 3) elicits a molecular response in cornea tissue. Lunar dust on the surface of the moon would have the added properties of ionization and activation potentially leading to further damage to the cornea and greater sensitivity to any other environmental insult such as exposure to radiation. Additional studies are required to fully assess the risk of vision impairment and the mechanistic responses initiated in cornea exposed to lunar dust as well as the potential for long-term effects to astronaut health

Optimization of immunostaining on flat-mounted human corneas.

PubMed

Forest, Fabien; Thuret, Gilles; Gain, Philippe; Dumollard, Jean-Marc; Peoc'h, Michel; Perrache, Chantal; He, Zhiguo

2015-01-01

In the literature, immunohistochemistry on cross sections is the main technique used to study protein expression in corneal endothelial cells (ECs), even though this method allows visualization of few ECs, without clear subcellular localization, and is subject to the staining artifacts frequently encountered at tissue borders. We previously proposed several protocols, using fixation in 0.5% paraformaldehyde (PFA) or in methanol, allowing immunostaining on flatmounted corneas for proteins of different cell compartments. In the present study, we further refined the technique by systematically assessing the effect of fixative temperature. Last, we used optimized protocols to further demonstrate the considerable advantages of immunostaining on flatmounted intact corneas: detection of rare cells in large fields of thousands of ECs and epithelial cells, and accurate subcellular localization of given proteins. The staining of four ubiquitous proteins, ZO-1, hnRNP L, actin, and histone H3, with clearly different subcellular localizations, was analyzed in ECs of organ-cultured corneas. Whole intact human corneas were fixed for 30 min in 0.5% paraformaldehyde or pure methanol at four temperatures (4 °C for PFA, -20 °C for methanol, and 23, 37, and 50 °C for both). Experiments were performed in duplicate and repeated on three corneas. Standardized pictures were analyzed independently by two experts. Second, optimized immunostaining protocols were applied to fresh corneas for three applications: identification of rare cells that express KI67 in the endothelium of specimens with Fuch's endothelial corneal dystrophy (FECD), the precise localization of neural cell adhesion molecules (NCAMs) in normal ECs and of the cytokeratin pair K3/12 and CD44 in normal epithelial cells, and the identification of cells that express S100b in the normal epithelium. Temperature strongly influenced immunostaining quality. There was no ubiquitous protocol, but nevertheless, room temperature may be recommended as first-line temperature during fixation, instead of the conventional -20 °C for methanol and 4 °C for PFA. Further optimization may be required for certain target proteins. Optimized protocols allowed description of two previously unknown findings: the presence of a few proliferating ECs in FECD specimens, suggesting ineffective compensatory mechanisms against premature EC death, and the localization of NCAMs exclusively in the lateral membranes of ECs, showing hexagonal organization at the apical pole and an irregular shape with increasing complexity toward the basal pole. Optimized protocols were also effective for the epithelium, allowing clear localization of cytokeratin 3/12 and CD44 in superficial and basal epithelial cells, respectively. Finally, S100b allowed identification of clusters of epithelial Langerhans cells near the limbus and more centrally. Fixative temperature is a crucial parameter in optimizing immunostaining on flatmounted intact corneas. Whole-tissue overview and precise subcellular staining are significant advantages over conventional immunohistochemistry (IHC) on cross sections. This technique, initially developed for the corneal endothelium, proved equally suitable for the corneal epithelium and could be used for other superficial mono- and multilayered epithelia.

White-Light, Dispersed-Fringe Interferometric Keratometer

NASA Technical Reports Server (NTRS)

Hochberg, Eric B.; Baroth, Edmund C.

1992-01-01

Proposed keratometer based on scheme involving spectral dispersal of white-light interference fringes. Instrument operates in "snapshot" mode: no scanning necessary, not necessary to immobilize patient's eye. Insensitive to vibration, involves no phase shifting, and has variable sensitivity. Intended primarily for use in medical assessments of human corneas, also used to measure shapes of animal corneas, lenses, and other aspherical or spherical reflective or partly reflective surfaces.

Pathogenesis of Acute and Delayed Corneal Lesions After Ocular Exposure to Sulfur Mustard Vapor

DTIC Science & Technology

2012-03-01

using a vapor cup delivery system. The transition from acute to delayed injury was characterized by clinical, histological, and ultrastructural metrics...These data demonstrate a system-based approach combining ultrastructural analysis , histochemistry, and molecular evaluation that links architectural...predictive of the 11% of corneas that underwent asymptomatic recovery. Ultrastructural comparison of asymptomatic and MGK corneas at 8 weeks indicates that MGK

Characterizing a Rat Brca2 Knockout Model

DTIC Science & Technology

2007-05-01

synechiae (adhesion of iris to lens), and anterior segment dysgenesis (adhesion of lens to cornea). The cataractous lens cells extend circumferentially...around the lens capsule to the posterior (arrowheads). ac, anterior chamber; c, cornea; i, iris ; l, lens; lc, lens capsule; r, retina; m, mineralization...to DSB forma - tion, chromosomal damage, and cell death resulting from ionizing radiation (reviewed by Pellegrini and Venkitaraman, 2004). Evidence from

Eye drop delivery of nano-polymeric micelle formulated genes with cornea-specific promoters.

PubMed

Tong, Yaw-Chong; Chang, Shwu-Fen; Liu, Chia-Yang; Kao, Winston W-Y; Huang, Chong Heng; Liaw, Jiahorng

2007-11-01

This study evaluates the eye drop delivery of genes with cornea-specific promoters, i.e., keratin 12 (K12) and keratocan (Kera3.2) promoters, by non-ionic poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) polymeric micelles (PM) to mouse and rabbit eyes, and investigates the underlying mechanisms. Three PM-formulated plasmids (pCMV-Lac Z, pK12-Lac Z and pKera3.2-Lac Z) containing the Lac Z gene for beta-galactosidase (beta-Gal) whose expression was driven by the promoter of either the cytomegalovirus early gene, the keratin 12 gene or the keratocan gene, were characterized by critical micelle concentration (CMC), dynamic light scattering (DLS), and atomic force microscopy (AFM). Transgene expression in ocular tissue after gene delivery was analyzed by 5-bromo-4-chloro-3-indolyl-beta-D-galactoside (X-Gal) color staining, 1,2-dioxetane beta-Gal enzymatic activity measurement, and real-time polymerase chain reaction (PCR) analysis. The delivery mechanisms of plasmid-PM on mouse and rabbit corneas were evaluated by EDTA and RGD (arginine-glycine-aspartic acid) peptide. The sizes of the three plasmid-PM complexes were around 150-200 nm with unimodal distribution. Enhanced stability was found for three plasmid-PM formulations after DNase I treatment. After six doses of eye drop delivery of pK12-Lac Z-PM three times a day, beta-Gal activity was significantly increased in both mouse and rabbit corneas. Stroma-specific Lac Z expression was only found in pKera3.2-Lac Z-PM-treated animals with pretreatment by 5 mM EDTA, an opener of junctions. Lac Z gene expression in both pK12-Lac Z-PM and pKera3.2-Lac Z-PM delivery groups was decreased by RGD peptide pretreatment. Cornea epithelium- and stroma-specific gene expression could be achieved using cornea-specific promoters of keratin 12 and keratocan genes, and the gene was delivered with PM formulation through non-invasive, eye drop in mice and rabbits. The transfection mechanism of plasmid-PM may involve endocytosis and particle size dependent paracellular transport. 2007 John Wiley & Sons, Ltd

Compensation of Corneal Oblique Astigmatism by Internal Optics: a Theoretical Analysis

PubMed Central

Liu, Tao; Thibos, Larry N.

2017-01-01

Purpose Oblique astigmatism is a prominent optical aberration of peripheral vision caused by oblique incidence of rays striking the refracting surfaces of the cornea and crystalline lens. We inquired whether oblique astigmatism from these two sources should be expected, theoretically, to have the same or opposite signs across the visual field at various states of accommodation. Methods Oblique astigmatism was computed across the central visual field for a rotationally-symmetric schematic-eye using optical design software. Accommodative state was varied by altering the apical radius of curvature and separation of the biconvex lens’s two aspheric surfaces in a manner consistent with published biometry. Oblique astigmatism was evaluated separately for the whole eye, the cornea, and the isolated lens over a wide range of surface curvatures and asphericity values associated with the accommodating lens. We also computed internal oblique astigmatism by subtracting corneal oblique astigmatism from whole-eye oblique astigmatism. Results A visual field map of oblique astigmatism for the cornea in the Navarro model follows the classic, textbook description of radially-oriented axes everywhere in the field. Despite large changes in surface properties during accommodation, intrinsic astigmatism of the isolated human lens for collimated light is also radially oriented and nearly independent of accommodation both in theory and in real eyes. However, the magnitude of ocular oblique astigmatism is smaller than that of the cornea alone, indicating partial compensation by the internal optics. This implies internal oblique astigmatism (which includes wavefront propagation from the posterior surface of the cornea to the anterior surface of the lens and intrinsic lens astigmatism) must have tangentially-oriented axes. This non-classical pattern of tangential axes for internal astigmatism was traced to the influence of corneal power on the angles of incidence of rays striking the internal lens. Conclusions Partial compensation of corneal astigmatism by internal optics is due mainly to the highly converging nature of wavefronts incident upon the lens resulting from corneal refraction. The degree of compensation is quadratically dependent on eccentricity but is expected to diminish as the eye accommodates. Neutralising the cornea by index-matching defeats internal compensation, revealing classical, radially-oriented oblique astigmatism in the isolated lens. PMID:28281302

Intrastromal corneal reshaping using a high-intensity femtosecond laser: A novel method of vision correction

NASA Astrophysics Data System (ADS)

Han, Taehee

A new technology to perform a minimally invasive cornea reshaping procedure has been developed. This can eliminate the incidence of the flap-related complications of the conventional eye refractive procedures by multiphoton processes using a very high-intensity (I ≥ 1013 W/cm 2), but low energy (Ep ˜ 100-200 microJ) femtosecond laser pulses. Due to much lower energy than that of the nanosecond laser pulses for the thermal photoablation, the multiphoton processes cause almost no collateral damage by heat and shock wave generation. In this method, a series of femtosecond laser pulses is used to create very narrow (< 30 microm) and sufficiently long (≥ 2.5 mm) micro-channels in the cornea. The micro-channels are oriented almost perpendicular to the eye's optical axis. Once the micro-channel reaches a desired length, another series of femtosecond pulses with higher intensity is efficiently delivered through the micro-channel to the endpoint where a certain amount of the stromal tissue is disintegrated by the multiphoton processes. The disintegrated fragments are ejected out of the cornea via the same micro-channel, allowing the corneal surface to collapse, and changing its refractive power. This new corneal reshaping method obviates any process of damaging the corneal surface layer, while retaining the advantages of the conventional refractive procedures such as Laser in situ keratomileusis (LASIK) and Photorefractive keratectomy (PRK). In order to demonstrate the flapless cornea reshaping procedure, we have conducted ex-vivo experiments on fresh porcine eyes. The reshaped corneas were evaluated by using optical coherence tomography (OCT). The test results have shown that this flapless intrastromal procedure can reshape the cornea as intended with almost no surface damage. We have also performed a series of experiments to demonstrate the multiphoton processes in the corneal tissue by very high-intensity femtosecond laser pulses. Through the optical emission spectroscopy, we investigated the spectral lines of calcium atom and ions from the femtosecond laser-induced plasma from the porcine corneal tissue. The experimental results have shown the intensity-dependence of ablation rate, which qualitatively verifies the characteristics of the multiphoton processes.

Global Survey of Corneal Transplantation and Eye Banking.

PubMed

Gain, Philippe; Jullienne, Rémy; He, Zhiguo; Aldossary, Mansour; Acquart, Sophie; Cognasse, Fabrice; Thuret, Gilles

2016-02-01

Corneal transplantation restores visual function when visual impairment caused by a corneal disease becomes too severe. It is considered the world's most frequent type of transplantation, but, to our knowledge, there are no exhaustive data allowing measurement of supply and demand, although such data are essential in defining local, national, and global strategies to fight corneal blindness. To describe the worldwide situation of corneal transplantation supply and demand. Data were collected between August 2012 and August 2013 from a systematic review of published literature in parallel with national and international reports on corneal transplantation and eye banking. In a second step, eye bank staff and/or corneal surgeons were interviewed on their local activities. Interviews were performed during international ophthalmology or eye-banking congresses or by telephone or email. Countries' national supply/demand status was classified using a 7-grade system. Data were collected from 148 countries. Corneal transplantation and corneal procurements per capita in each country. In 2012, we identified 184,576 corneal transplants performed in 116 countries. These were procured from 283,530 corneas and stored in 742 eye banks. The top indications were Fuchs dystrophy (39% of all corneal transplants performed), a primary corneal edema mostly affecting elderly individuals; keratoconus (27%), a corneal disease that slowly deforms the cornea in young people; and sequellae of infectious keratitis (20%). The United States, with 199.10-6 corneal transplants per capita, had the highest transplantation rate, followed by Lebanon (122.10-6) and Canada (117.10-6), while the median of the 116 transplanting countries was 19.10-6. Corneas were procured in only 82 countries. Only the United States and Sri Lanka exported large numbers of donor corneas. About 53% of the world's population had no access to corneal transplantation. Our survey globally quantified the considerable shortage of corneal graft tissue, with only 1 cornea available for 70 needed. Efforts to encourage cornea donation must continue in all countries, but it is also essential to develop alternative and/or complementary solutions, such as corneal bioengineering.

Effects of immersion solution on corneal tissue strip hydration and thickness during thermal shrinkage

NASA Astrophysics Data System (ADS)

Borja, David; Manns, Fabrice; Lamar, Peggy D.; Rosen, Alexander; Parel, Jean-Marie A.

2003-07-01

Purpose: To assess the effects of immersion solutions with different Dextran concentrations on the hydration of cornea tissue strips at normal body temperature. Methods: A 20% Dextran-BSS solution was injected via a self sealing limbal-transcorneal tunnel incision using a 30ga needle into the anterior chamber of human donor eyes until the globe was hard. The eyes were then immersed cornea down overnight in the same solution. Corneal thickness was measured by ultrasound pachymetry after the eyes were re-inflated and at regular intervals to assess dehydration. When the central cornea thickness reached 400-500μm corneal buttons were removed using a 10mm trephine. The buttons were then cut into 6x6mm strips using a custom-made jig and immediately immersed in solutions of Dextran (15 to 20% in increments of 2.5%) at 35degC. The edge thickness of the immersed strip was measured every 5 min for one hour using an optical comparator (Topcon, Japan) modified for tissue shadowphotogrammetry. Results: For five Florida Lions Eye Bank donated eyes after one hour in the Dextran solution the mean final measured thickness of corneas in 20%, 17.5% and 15% Dextran-BSS solutions were 570 (+/-75) μm, 680 (+/-70) μm, and 1080 (+/-95) μm respectively. These measured thicknesses changes correspond to an average swelling of 1.2, 1.4 and 2.2 times the initial thickness of each cornea strip in the 20%, 17.5% and 15% Dextran-BSS solutions respectively. Conclusion: We will conduct experiments at higher Dextran concentrations but our current results indicate that 20% Dextran-BSS solution was the best solution tested at keeping corneal tissue strips at or close to physiological thickness.

Cornea lymphatics drive the CD8+ T-cell response to herpes simplex virus-1.

PubMed

Gurung, Hem R; Carr, Meghan M; Carr, Daniel J J

2017-01-01

Herpes simplex virus-1 (HSV-1) infection of the cornea induces vascular endothelial growth factor A (VEGF-A)-dependent lymphangiogenesis. However, the extent to which HSV-1-induced corneal lymphangiogenesis impacts the adaptive immune response has not been characterized. Here, we used floxed VEGF-A mice to study the importance of newly created corneal lymphatic vessels in the host adaptive immune response to infection. Whereas the mice infected with the parental virus (strain SC16) exhibited robust corneal lymphangiogenesis, mice that received the recombinant virus (SC16 ICP0-Cre) that expresses Cre recombinase under the control of infected cell protein 0 (ICP0), an HSV-1 immediate-early gene, showed a significant reduction in lymphangiogenesis. There was no difference in virus recovered from the cornea of mice infected with SC16 vs SC16 ICP0-Cre. However, viral loads were significantly elevated in the trigeminal ganglia (TG) of mice with reduced corneal lymphangiogenesis. The increase in viral titer correlated with a significant loss of HSV-1-specific CD8 + T cells that traffic to the TG of mice infected with the recombinant virus. Intrastromal delivery of size-exclusion dye (fluorescein isothiocyanate-dextran) revealed a time-dependent defect in the ability of the lymphatic vessels in SC16 ICP0-Cre-infected mice to transport soluble antigen from the cornea to the draining lymph nodes. We interpret these results to suggest that the newly created lymphatic vessels in the cornea driven by HSV-1 infection are critical in the delivery of soluble viral antigen to the draining lymph node and subsequent development of the CD8 + T-cell response to HSV-1.

Controlled reshaping of the front surface of the cornea through its full-area ablation outside of the optical zone with a Gaussian ArF excimer laser beam

NASA Astrophysics Data System (ADS)

Semchishen, A. V.; Semchishen, V. A.

2014-01-01

We studied in vitro the response of the topography of the cornea to its full-area laser ablation (the laser beam spot diameter is commensurable with the size of the interface) outside of the central zone with an excimer laser having a Gaussian fluence distribution across the beam. Subject to investigation were the topographically controlled surface changes of the anterior cornea in 60 porcine eyes with a 5 ± 1.25-diopter artificially induced astigmatism, the changes being caused by laser ablation of the stromal collagen in two 3.5-mm-dia. circular areas along the weaker astigmatism axis. Experimental relationships are presented between the actual astigmatism correction and the expected correction for the intact optical zones 1, 2, 3, and 4 mm in diameter. The data for each zone were approximated by the least-squares method with the function d = a + bx. The coefficient b is given with the root-mean-square error. The statistical processing of the data yielded the following results: d = (0.14 ± 0.037)x for the 1-mm-dia. optical zone, (1.10 ± 0.036)x for the 2-mm-dia. optical zone, (1.04 ± 0.020)x for the 3-mm-dia. optical zone, and (0.55 ± 0.04)x for the 4-mm-dia. optical zone. Full astigmatism correction was achieved with ablation effected outside of the 3-mm-dia. optical zone. The surface changes of the cornea are shown to be due not only to the removal of the corneal tissue, but also to the biomechanical topographic response of the cornea to its strain caused by the formation of a dense pseudomembrane in the ablation area.

Quantitative 3-D Corneal Imaging In Vivo Using a Modified HRT- RCM Confocal Microscope

PubMed Central

Petroll, W. Matthew.; Weaver, Matthew; Vaidya, Saurabh; McCulley, James P.; Cavanagh, H. Dwight

2012-01-01

Purpose The purpose of this study was to develop and test hardware and software modifications to allow quantitative full-thickness corneal imaging using the HRT Rostock Corneal Module. Methods A PC-controlled motor drive with positional feedback was integrated into the system to allow automated focusing through the entire cornea. The left eyes of ten New Zealand White rabbits were scanned from endothelium to epithelium. Image sequences were read into a custom-developed program for depth calculation and measurement of sub-layer thicknesses. 3-D visualizations were also generated using Imaris. In six rabbits, stack images were registered, and depth-dependent counts of keratocyte nuclei were made using Metamorph. Results The mean epithelial and corneal thicknesses measured in the rabbit were 47 ± 5 μm and 373 ± 25 μm, respectively (N = 10 corneas); coefficients of variation for repeated scans were 8.2% and 2.1%. Corneal thickness measured using ultrasonic pachymetry was 374 ± 17 μm. The mean overall keratocyte density measured in the rabbit was 43,246 ± 5,603 cells/mm3 in vivo (N = 6 corneas). There was a gradual decrease in keratocyte density from the anterior to posterior cornea (R = 0.99), consistent with previous data generated in vitro. Conclusions This modified system allows high resolution 3-D image stacks to be collected from the full thickness rabbit cornea in vivo. These datasets can be used for interactive visualization of corneal cell layers, measurement of sub-layer thickness, and depth-dependent keratocyte density measurements. Overall, the modifications significantly expand the potential quantitative research applications of the HRT-RCM microscope. PMID:23051907

Why Does the Healthy Cornea Resist Pseudomonas aeruginosa Infection?

PubMed Central

Evans, David J.; Fleiszig, Suzanne M. J.

2013-01-01

Purpose To provide our perspective on why the cornea is resistant to infection based on our research results with Pseudomonas aeruginosa. Perspective We focus on our current understanding of the interplay between bacteria, tear fluid and the corneal epithelium that determine health as the usual outcome, and propose a theoretical model for how contact lens wear might change those interactions to enable susceptibility to P. aeruginosa infection. Methods Use of “null-infection” in vivo models, cultured human corneal epithelial cells, contact lens-wearing animal models, and bacterial genetics help to elucidate mechanisms by which P. aeruginosa survive at the ocular surface, adheres, and traverses multilayered corneal epithelia. These models also help elucidate the molecular mechanisms of corneal epithelial innate defense. Results and Discussion Tear fluid and the corneal epithelium combine to make a formidable defense against P. aeruginosa infection of the cornea. Part of that defense involves the expression of antimicrobials such as β-defensins, the cathelicidin LL-37, cytokeratin-derived antimicrobial peptides, and RNase7. Immunomodulators such as SP-D and ST2 also contribute. Innate defenses of the cornea depend in part on MyD88, a key adaptor protein of TLR and IL-1R signaling, but the basal lamina represents the final barrier to bacterial penetration. Overcoming these defenses involves P. aeruginosa adaptation, expression of the type three secretion system, proteases, and P. aeruginosa biofilm formation on contact lenses. Conclusion After more than two decades of research focused on understanding how contact lens wear predisposes to P. aeruginosa infection, our working hypothesis places blame for microbial keratitis on bacterial adaptation to ocular surface defenses, combined with changes to the biochemistry of the corneal surface caused by trapping bacteria and tear fluid against the cornea under the lens. PMID:23601656

Coenzyme Q10 instilled as eye drops on the cornea reaches the retina and protects retinal layers from apoptosis in a mouse model of kainate-induced retinal damage.

PubMed

Lulli, Matteo; Witort, Ewa; Papucci, Laura; Torre, Eugenio; Schipani, Christian; Bergamini, Christian; Dal Monte, Massimo; Capaccioli, Sergio

2012-12-17

To evaluate if coenzyme Q10 (CoQ10) can protect retinal ganglion cells (RGCs) from apoptosis and, when instilled as eye drops on the cornea, if it can reach the retina and exert its antiapoptotic activity in this area in a mouse model of kainate (KA)-induced retinal damage. Rat primary or cultured RGCs were subjected to glutamate (50 μM) or chemical hypoxia (Antimycin A, 200 μM) or serum withdrawal (FBS, 0.5%) in the presence or absence of CoQ10 (10 μM). Cell viability was evaluated by light microscopy and fluorescence-activated cell sorting analyses. Apoptosis was evaluated by caspase 3/7 activity and mitochondrion depolarization tetramethylrhodamine ethyl ester analysis. CoQ10 transfer to the retina following its instillation as eye drops on the cornea was quantified by HPLC. Retinal protection by CoQ10 (10 μM) eye drops instilled on the cornea was then evaluated in a mouse model of KA-induced excitotoxic retinal cell apoptosis by cleaved caspase 3 immunohistofluorescence, caspase 3/7 activity assays, and quantification of inhibition of RGC loss. CoQ10 significantly increased viable cells by preventing RGC apoptosis. Furthermore, when topically applied as eye drops to the cornea, it reached the retina, thus substantially increasing local CoQ10 concentration and protecting retinal layers from apoptosis. The ability of CoQ10 eye drops to protect retinal cells from apoptosis in the mouse model of KA-induced retinal damage suggests that topical CoQ10 may be evaluated in designing therapies for treating apoptosis-driven retinopathies.

Expression profiles of aquaporins in rat conjunctiva, cornea, lacrimal gland and Meibomian gland.

PubMed

Yu, Dongfang; Thelin, William R; Randell, Scott H; Boucher, Richard C

2012-10-01

The aim of the study was to elucidate aquaporin (AQP) family member mRNA expression and protein expression/localization in the rat lacrimal functional unit. The mRNA expression of all rat AQPs (AQP0-9, 11-12) in palpebral, fornical, and bulbar conjunctiva, cornea, lacrimal gland, and Meibomian gland was measured by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and real time RT-PCR. Antibodies against AQP1, 3, 4, 5, 9, and 11 were used in Western blotting and immunohistochemistry to determine protein expression and distribution. Our study demonstrated characteristic AQP expression profiles in rat ocular tissues. AQP1, 3, 4, 5, 8, 9, 11, and 12 mRNA were detected in conjunctiva. AQP0, 1, 2, 3, 4, 5, 6, 11, and 12 mRNA were expressed in cornea. AQP0, 1, 2, 3, 4, 5, 7, 8, and 11 mRNA were detected in lacrimal gland. AQP1, 3, 4, 5, 7, 8, 9, 11, and 12 mRNA were identified in Meibomian gland. By Western blot, AQP1, 3, 5, and 11 were detected in conjunctiva; AQP1, 3, 5, and 11 were identified in cornea; AQP1, 3, 4, 5, and 11 were detected in lacrimal gland; and AQP1, 3, 4, 5, 9, and 11 were present in Meibomian gland. Immunohistochemistry localized AQPs to distinct sites in the various tissues. This study rigorously analyzed AQPs expression and localization in rat conjunctiva, cornea, lacrimal gland, and Meibomian gland tissues. Our findings provide a comprehensive platform for further investigation into the physiological or pathophysiological relevance of AQPs in ocular surface. Copyright © 2012 Elsevier Ltd. All rights reserved.

The effect of standard and high-fluence corneal cross-linking (CXL) on cornea and limbus.

PubMed

Richoz, Olivier; Tabibian, David; Hammer, Arthur; Majo, François; Nicolas, Michael; Hafezi, Farhad

2014-07-22

When treating peripheral ectatic disease-like pellucid marginal degeneration (PMD), corneal cross-linking with UV-A and riboflavin (CXL) must be applied eccentrically to the periphery of the lower cornea, partly irradiating the corneal limbus. Here, we investigated the effect of standard and double-standard fluence corneal cross-linking with riboflavin and UV-A (CXL) on cornea and corneal limbus in the rabbit eye in vivo. Epithelium-off CXL was performed in male New Zealand White rabbits with two irradiation diameters (7 mm central cornea, 13 mm cornea and limbus), using standard fluence (5.4 J/cm(2)) and double-standard fluence (10.8 J/cm(2)) settings. Controls were subjected to epithelial removal and riboflavin instillation, but were not irradiated with UV-A. Following CXL, animals were examined daily until complete closure of the epithelium, and at 7, 14, 21, and 28 days. Animals were killed and a corneoscleral button was excised and processed for light microscopy and immunohistochemistry. For both irradiation diameters and fluences tested, no signs of endothelial damage or limbal vessel thrombosis were observed, and time to re-epithelialization was similar to untreated controls. Histological and immunohistochemical analysis revealed no differences in the p63 putative stem cell marker expression pattern. Even when using fluence twice as high as the one used in current clinical CXL settings, circumferential UV-A irradiation of the corneal limbus does not alter the regenerative capacity of the limbal epithelial cells, and the expression pattern of the putative stem cell marker p63 remains unchanged. This suggests that eccentric CXL may be performed safely in PMD. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

Corneal NF-kappaB activity is necessary for the retention of transparency in the cornea of UV-B-exposed transgenic reporter mice.

PubMed

Alexander, George; Carlsen, Harald; Blomhoff, Rune

2006-04-01

To determine the dynamics of Nuclear Factor-kappaB (NF-kappaB) in murine corneal pathology and the role of NF-kappaB in maintaining corneal clarity after ultraviolet B radiation insult, transgenic mice containing NF-kappaB-luciferase reporter were exposed to LPS (bacterial lipopolysaccharide), TNF-alpha (Tumor Necrosis Factor-alpha) or 4 kJ m(-2) UV-B radiation. NF-kappaB decoy oligonucleotides were also administered in some of the UV-B experiments. Following various exposure times, the mice were sacrificed and whole eyes or corneal tissues were obtained. Whole eyes were examined for scattering using a point-source optical imaging technique. Tissue homogenates were examined for luciferase activity using a luminometer. TNF-alpha and LPS-injected NF-kappaB-luciferase transgenic mice demonstrated 3-10-fold increases in cornea NF-kappaB with peak activities at 4 and 6 hr post-injection, respectively. Mice exposed to 4 kJ m(-2) UV-B exhibited a 3-fold increase in NF-kappaB activity 4 hr post-exposure. The administration of NF-kappaB-decoy oligonucleotides to mice had the effect of reducing UV-B-induced NF-kappaB activity in the cornea and significantly increasing the amount of light scattering in UV-B exposed corneas 7 days post-UV-B exposure when compared to sham injected mice. These results indicate that NF-kappaB is activated in cornea in pathologies that involves increased plasma levels of LPS and TNF-alpha, as well as direct UV-B exposure, and suggest that NF-kappaB activation play an essential part in the corneal healing process.

Characterization of Ocular Biomechanics in Pellucid Marginal Degeneration.

PubMed

Lenk, Janine; Haustein, Michael; Terai, Naim; Spoerl, Eberhard; Raiskup, Frederik

2016-04-01

This study sought to investigate the diagnostic capacity of corneal biomechanical response parameters in a group of patients with pellucid marginal degeneration (PMD) using the Ocular Response Analyzer (ORA) and Corvis ST devices. In this prospective clinical study, we used the Corvis ST and ORA devices to investigate the ocular biomechanics of patients with PMD. Eighty-one eyes were included, and 2 study groups were formed: the PMD group (the study group, n = 29) and the control group (n = 52). We focused on 13 biomechanical parameters. Statistical analysis was performed using SPSS. Biomechanical parameters for the 2 groups were compared using analysis of covariance. The ORA results demonstrated that the Keratoconus Match Index was significantly lower in the PMD group than in the control group (0.031 ± 0.37 vs. 0.79 ± 0.33; P = 0.001). The 2 groups did not significantly differ with respect to intraocular pressure- and central corneal thickness-adjusted values for corneal hysteresis or corneal resistance factor. Regarding the Corvis parameters, differences between the control and PMD groups were detected for CorWmax amp (control 1.01 ± 0.01, PMD 1.06 ± 0.01; P = 0.020) and CorA2 t (control 21.78 ± 0.03, PMD 21.66 ± 0.04; P = 0.0003). We identified 2 Corvis parameters that could be used to characterize PMD and differentiate PMD corneas from normal corneas. These parameters support the hypothesis that there is significantly less deformation of the central cornea in PMD corneas than in healthy corneas. However, because useful "first-line" diagnostic devices for diagnosing PMD (such as Pentacam and the ORA) exist, the Corvis ST serves as an additional diagnostic tool that can also be used for long-term monitoring after diagnosis confirmation.

Corneal densitometry and its correlation with age, pachymetry, corneal curvature, and refraction.

PubMed

Garzón, Nuria; Poyales, Francisco; Illarramendi, Igor; Mendicute, Javier; Jáñez, Óscar; Caro, Pedro; López, Alfredo; Argüeso, Francisco

2017-12-01

To determine normative corneal densitometry values in relation to age, sex, refractive error, corneal thickness, and keratometry, measured using the Oculus Pentacam system. Three hundred and thirty-eight healthy subjects (185 men; 153 women) with no corneal disease underwent an exhaustive ocular examination. Corneal densitometry was expressed in standardized grayscale units (GSU). The mean corneal densitometry over the total area was 16.46 ± 1.85 GSU. The Pearson correlation coefficient for total densitometry was r = 0.542 (p < 0.001). Statistically significant differences were found between men and women for the total area (p = 0.006), with readings of 16.22 ± 1.54 GSU and 16.60 ± 1.83 GSU, respectively. When the cornea was divided into layers of different depths, a significant correlation was found for all layers and age: r = 0.447 (p < 0.001), r = 0.563 (p < 0.001), and r = 0.520 (p < 0.001) for the anterior, central, and posterior layers, respectively. However, when the cornea was divided into concentric annuli starting from the center of the cornea, densitometry was strongly correlated only with age in the 6-10-mm annulus (p < 0.001). Neither mean keratometry nor spherical equivalent was correlated with corneal densitometry in any zone of the cornea (p > 0.05). This is the first report of normative corneal densitometry values in relation to keratometry, corneal thickness, and spherical equivalent measured with the latest Oculus Pentacam software. Corneal densitometry increases with age, but corneal keratometry and refractive parameters do not affect light scattering in the human cornea.

Number of Bacteria and Time of Coincubation With Bacteria Required for the Development of Acanthamoeba Keratitis.

PubMed

Nakagawa, Hayate; Hattori, Takaaki; Koike, Naohito; Ehara, Tomoko; Narimatsu, Akitomo; Kumakura, Shigeto; Matsumoto, Tetsuya; Goto, Hiroshi

2017-03-01

We hypothesized that bacteria may be a factor contributing to the development of Acanthamoeba keratitis (AK). We investigated interactions between Acanthamoeba and Pseudomonas aeruginosa for the development of keratitis in rabbit corneas. Acanthamoeba castellanii (ATCC50492) and P. aeruginosa (PAO-1) were used. Two densities of P. aeruginosa (high, 1 × 10/mL; low, 3 × 10/mL) and 2 durations of coincubation (long, 6 h; short, 2 h) of Acanthamoeba with 1 × 10/mL of P. aeruginosa were tested. Acanthamoeba alone or Acanthamoeba coincubated with P. aeruginosa was inoculated into rabbit corneas. After inoculation, levofloxacin (LVFX) eye drops were administered. The clinical score of the cornea was evaluated after inoculation. Acanthamoeba alone did not produce keratitis during a 5-day observation period. Rabbit corneas inoculated with Acanthamoeba coincubated with low-density P. aeruginosa followed by topical LVFX were clear with few infiltrates. Corneas inoculated with Acanthamoeba coincubated with high-density P. aeruginosa followed by LVFX treatment developed severe keratitis, and clinical scores were significantly higher compared with high-density P. aeruginosa alone followed by LVFX treatment (scores 7, 9.6, 8.5 vs. 3, 3.5, 3.25 on days 1-3, all P < 0.01). The long (6 h) coincubation time of Acanthamoeba with high-density P. aeruginosa resulted in more severe keratitis compared with short (2 h) coincubation (scores, 9.7, 12.7, 12.1, 9.8, 8.7 vs. 7, 9.6, 8.5, 6.9, 5.6 on days 1-5, all P < 0.01). These results suggest that the presence of bacteria is essential and a critical number of bacteria is required for the development of AK. The time of coexistence with bacteria may be an important determinant of the severity of AK.

Frequency of Complications During Preparation of Corneal Lamellae Used in Posterior Lamellar Keratoplasty Using the Pneumodissection Technique (Big Bubble).

PubMed

Studeny, Pavel; Netukova, Magdalena; Hlozanek, Martin; Bednar, Jan; Jirsova, Katerina; Krizova, Deli

2018-04-26

To determine the frequency of formation of various types of bubbles and the potential impact of donor and lamella parameters on this frequency, and to identify possible risk factors of unsuccessful "big-bubble" creation in preparation of pre-Descemet endothelial keratoplasty and Descemet membrane endothelial keratoplasty with peripheral stromal support. Donor age and sex, death to preservation time (DPT), storage time, presence of corneal scars (mainly a condition after cataract surgery), and endothelial cell density of 256 donor corneas were assessed before Descemet membrane endothelial keratoplasty with peripheral stromal support or pre-Descemet endothelial keratoplasty lamella preparation using the big-bubble technique. Mean donor age was 62.3 ± 8.5 years (28.3% women and 71.7% men). Mean endothelial cell density of the donor graft was 2866 ± 255 cells/mm. Mean DPT was 10.12 ± 4.88 hours, and mean storage time of the transplant before surgery was 6.5 ± 4.8 days. Corneal scars were present in 17 donor grafts (6.6%) after cataract surgery. Eleven corneas were devalued because of Descemet membrane rupture during preparation (4.3%). In 182 corneas, standard bubble type I was created (71.7%); in 27 corneas, bubble type II was created; eventually, both types of bubbles formed simultaneously (10.5%); in 47 corneas, no bubble was created (18.4%). We identified higher endothelial cell density, shorter DPT, and the presence of corneal scars after cataract surgery as risk factors threatening successful bubble formation. The only risk factor for creating type II bubbles was higher donor age in our study.

Revisited microanatomy of the corneal endothelial periphery: new evidence for continuous centripetal migration of endothelial cells in humans.

PubMed

He, Zhiguo; Campolmi, Nelly; Gain, Philippe; Ha Thi, Binh Minh; Dumollard, Jean-Marc; Duband, Sébastien; Peoc'h, Michel; Piselli, Simone; Garraud, Olivier; Thuret, Gilles

2012-11-01

The control of corneal transparency depends on the integrity of its endothelial monolayer, which is considered nonregenerative in adult humans. In pathological situations, endothelial cell (EC) loss, not offset by mitosis, can lead to irreversible corneal edema and blindness. However, the hypothesis of a slow, clinically insufficient regeneration starting from the corneal periphery remains debatable. The authors have re-evaluated the microanatomy of the endothelium in order to identify structures likely to support this homeostasis model. Whole endothelia of 88 human corneas (not stored, and stored in organ culture) with mean donor age of 80 ± 12 years were analyzed using an original flat-mounting technique. In 61% of corneas, cells located at the extreme periphery (last 200 μm of the endothelium) were organized in small clusters with two to three cell layers around Hassall-Henle bodies. In 68% of corneas, peripheral ECs formed centripetal rows 830 ± 295 μm long, with Descemet membrane furrows visible by scanning electron microscopy. EC density was significantly higher in zones with cell rows. When immunostained, ECs in the extreme periphery exhibited lesser differentiation (ZO-1, Actin, Na/K ATPase, CoxIV) than ECs in the center of the cornea but preferentially expressed stem cell markers (Nestin, Telomerase, and occasionally breast cancer resistance protein) and, in rare cases, the proliferation marker Ki67. Stored corneas had fewer cell clusters but more Ki67-positive ECs. We identified a novel anatomic organization in the periphery of the human corneal endothelium, suggesting a continuous slow centripetal migration, throughout life, of ECs from specific niches. Copyright © 2012 AlphaMed Press.

Alterations in Corneal Sensory Nerves During Homeostasis, Aging, and After Injury in Mice Lacking the Heparan Sulfate Proteoglycan Syndecan-1.

PubMed

Pal-Ghosh, Sonali; Tadvalkar, Gauri; Stepp, Mary Ann

2017-10-01

To determine the impact of the loss of syndecan 1 (SDC1) on intraepithelial corneal nerves (ICNs) during homeostasis, aging, and in response to 1.5-mm trephine and debridement injury. Whole-mount corneas are used to quantify ICN density and thickness over time after birth and in response to injury in SDC1-null and wild-type (WT) mice. High-resolution three-dimensional imaging is used to visualize intraepithelial nerve terminals (INTs), axon fragments, and lysosomes in corneal epithelial cells using antibodies against growth associated protein 43 (GAP43), βIII tubulin, and LAMP1. Quantitative PCR was performed to quantify expression of SDC1, SDC2, SDC3, and SDC4 in corneal epithelial mRNA. Phagocytosis was assessed by quantifying internalization of fluorescently labeled 1-μm latex beads. Intraepithelial corneal nerves innervate the corneas of SDC1-null mice more slowly. At 8 weeks, ICN density is less but thickness is greater. Apically projecting intraepithelial nerve terminals and lysosome-associated membrane glycoprotein 1 (LAMP1) are also reduced in unwounded SDC1-null corneas. Quantitative PCR and immunofluorescence studies show that SDC3 expression and localization are increased in SDC1-null ICNs. Wild-type and SDC1-null corneas lose ICN density and thickness as they age. Recovery of axon density and thickness after trephine but not debridement wounds is slower in SDC1-null corneas compared with WT. Experiments assessing phagocytosis show reduced bead internalization by SDC1-null epithelial cells. Syndecan-1 deficiency alters ICN morphology and homeostasis during aging, reduces epithelial phagocytosis, and impairs reinnervation after trephine but not debridement injury. These data provide insight into the mechanisms used by sensory nerves to reinnervate after injury.

MRP8/14 Enhances Corneal Susceptibility to Pseudomonas aeruginosa Infection by Amplifying Inflammatory Responses

PubMed Central

Deng, Qiuchan; Sun, Mingxia; Yang, Kun; Zhu, Min; Chen, Kang; Yuan, Jin; Wu, Minhao; Huang, Xi

2013-01-01

Purpose. We explored the role of myeloid-related protein 8 and 14 (MRP8/14) in Pseudomonas aeruginosa (PA) keratitis. Methods. MRP8/14 mRNA levels in human corneal scrapes and mouse corneas infected by PA were tested using real-time PCR. MRP8/14 protein expression in C57BL/6 (B6) corneas was confirmed using Western blot assay and immunohistochemistry. B6 mice were injected subconjunctivally with siRNA for MRP8/14, and then infected with PA. Bacterial plate counts and myeloperoxidase assays were used to determine the bacterial load and polymorphonuclear neutrophil (PMN) infiltration in infected B6 corneas. Pro-inflammatory cytokine levels in vivo and in vitro were examined with PCR and ELISA. In murine macrophage-like RAW264.7 cells, phagocytosis and bacterial killing were assessed using plate count assays, and reactive oxygen species (ROS) and nitric oxide (NO) levels were tested with flow cytometry and Griess assay, respectively. Results. MRP8/14 expression levels were increased significantly in human corneal scrapes and B6 corneas after PA infection. Silencing of MRP8/14 in B6 corneas significantly reduced the severity of corneal disease, bacterial clearance, PMN infiltration, and pro-inflammatory cytokine expression after PA infection. In vitro studies demonstrated further that silencing of MRP8/14 suppressed pro-inflammatory cytokine production, bacterial killing, and ROS production, but not phagocytosis or NO production. Conclusions. Our study demonstrated a dual role for MRP8/14 in bacterial keratitis. Although MRP8/14 promotes bacterial clearance by enhancing ROS production, it functions more importantly as an inflammatory amplifier at the ocular surface by enhancing pro-inflammatory cytokine expression, thus contributing to the corneal susceptibility. PMID:23299480

SLC4A11 Prevents Osmotic Imbalance Leading to Corneal Endothelial Dystrophy, Deafness, and Polyuria*

PubMed Central

Gröger, Nicole; Fröhlich, Henning; Maier, Hannes; Olbrich, Andrea; Kostin, Sawa; Braun, Thomas; Boettger, Thomas

2010-01-01

Maintenance of ion concentration gradients is essential for the function of many organs, including the kidney, the cornea, and the inner ear. Ion concentrations and fluid content in the cornea are regulated by endothelial cells that separate the collagenous avascular corneal stroma from the anterior eye chamber. Failure to maintain correct ion concentrations leads to swelling and destruction of the cornea. In the inner ear, the stria vascularis is responsible for generating proper ion concentrations in the endolymph, which is essential for hearing. Mutations of SLC4A11 in humans lead to syndromes associated with corneal dystrophy and perceptive deafness. The molecular mechanisms underlying these symptoms are poorly understood, impeding therapeutic interventions. The ion transporter SLC4A11 mediates sodium-dependent transport of borate as well as flux of sodium and hydroxyl ions in vitro. Here, we show that SLC4A11 is expressed in the endothelial cells of the cornea where it prevents severe morphological changes of the cornea caused by increased sodium chloride concentrations in the stroma. In the inner ear, SLC4A11 is located in fibrocytes underlying the stria vascularis. Loss of SLC4A11 leads to morphological changes in the fibrocytes and deafness. We demonstrate that SLC4A11 is essential for the generation of the endocochlear potential but not for regulation of potassium concentrations in the endolymph. In the kidney, SLC4A11 is expressed in the thin descending limb of Henle loop. SLC4A11 is essential for urinary concentration, suggesting that SLC4A11 participates in the countercurrent multiplication that concentrates urine in the kidney medulla. PMID:20185830

Effects of topical nonsteroidal antiinflammatory drugs on the expression of matrix metalloproteinases in the cornea.

PubMed

Reviglio, Victor E; Rana, Tayyib S; Li, Qian J; Ashraf, M Farooq; Daly, Mary K; O'Brien, Terrence P

2003-05-01

To assess the effects of nonsteroidal antiinflammatory drug (NSAID) eyedrops on the expression of matrix metalloproteinases in corneal tissue. Ocular Microbiology and Immunology Laboratory, Refractive Surgery Research Laboratory, The Wilmer Eye Institute, Johns Hopkins University, Baltimore, Maryland, USA. Seventy rats were divided into 2 groups: intact and debrided epithelium. Uniform central corneal epithelial defects were created in the right eye of the debrided corneal group. Each group was divided into 4 subgroups, each receiving 1 of the following eyedrops or artificial tears: The 3 NSAIDs were diclofenac sodium 0.1% (Falcon or Voltaren) and preservative-free ketorolac 0.5% (Acular PF). The artificial tears were carboxymethylcellulose sodium 0.5% (Refresh Plus PF). The eyedrops were administered 4 times a day for 1 week. The rats were killed on days 2 and 7. The corneas were excised and processed for immunohistochemical staining, Western blot assay, and zymography studies to determine the localization of the production of the following matrix metalloproteinases (MMPs): MMP-1, MMP-2, MMP-8, and MMP-9. Matrix metalloproteinase-1, MMP-8, and MMP-2 were detected in rat corneas at 48 hours in the debrided and intact epithelium groups treated with NSAID eyedrops. The MMP-1 and MMP-8 expression levels were higher in intact corneas in the diclofenac sodium groups than in the ketorolac and artificial tears groups. The expression was localized mostly in the epithelial cells and occasionally in keratocytes. This study provides preliminary evidence that topical application of some NSAIDs can induce the early expression of MMP-1, MMP-2, and MMP-8 in the cornea, suggesting that MMPs play a role in the corneal cytotoxicity of certain NSAIDs.

Potentiation of femtosecond laser intratissue refractive index shaping (IRIS) in the living cornea with sodium fluorescein.

PubMed

Nagy, Lana J; Ding, Li; Xu, Lisen; Knox, Wayne H; Huxlin, Krystel R

2010-02-01

To assess the effectiveness of intratissue refractive index shaping (IRIS) in living corneas and test the hypothesis that it can be enhanced by increasing the two-photon absorption (TPA) of the tissue. Three corneas were removed from adult cats and cut into six pieces, which were placed in preservative (Optisol-GS; Bausch & Lomb, Inc., Irvine, CA) containing 0%, 0.25%, 1%, 1.5%, or 2.5% sodium fluorescein (Na-Fl). An 800-nm Ti:Sapphire femtosecond laser with a 100-fs pulse duration and 80-MHz repetition rate was used to perform IRIS in each piece, creating several refractive index (RI) modification lines at different speeds (between 0.1 and 5 mm/s). The lines were 1 mum wide, 10 microm apart, and approximately 150 microm below the tissue surface. The RI change of each grating was measured using calibrated, differential interference contrast microscopy. TUNEL staining was performed to assess whether IRIS or Na-Fl doping causes cell death. Scanning at 0.1 mm/s changed the RI of undoped, living corneas by 0.005. In doped corneas, RI changes between 0.01 and 0.02 were reliably achieved with higher scanning speeds. The magnitude of RI changes attained was directly proportional to Na-Fl doping concentration and inversely proportional to the scanning speed used to create the gratings. IRIS can be efficiently performed in living corneal tissue. Increasing the TPA of the tissue with Na-Fl increased both the scanning speeds and the magnitude of RI changes in a dose-dependent manner. Ongoing studies are exploring the use of IRIS to alter the optical properties of corneal tissue in situ, over an extended period.

Regional assessment of energy-producing metabolic activity in the endothelium of donor corneas.

PubMed

Greiner, Mark A; Burckart, Kimberlee A; Wagoner, Michael D; Schmidt, Gregory A; Reed, Cynthia R; Liaboe, Chase A; Flamme-Wiese, Miles J; Zimmerman, M Bridget; Mullins, Robert F; Kardon, Randy H; Goins, Kenneth M; Aldrich, Benjamin T

2015-05-01

We characterized mitochondrial respiration and glycolysis activity of human corneal endothelium, and compared metabolic activity between central and peripheral regions. Endothelial keratoplasty-suitable corneas were obtained from donors aged 50 to 75 years. The endothelium-Descemet membrane complex (EDM) was isolated, and 3-mm punches were obtained from central and peripheral regions. Endothelium-Descemet membrane punches were assayed for mitochondrial respiration (oxygen consumption) and glycolysis (extracellular acidification) using an extracellular flux analyzer. Enzymatic (citrate synthase, glucose hexokinase) and mitochondrial density (MitoTracker) assays also were performed. Ten corneas were analyzed per assay. Metabolic activity for mitochondrial respiration and glycolysis showed expected changes to assay compounds (P < 0.01, all pairwise comparisons). Basal mitochondrial respiration and glycolysis activity did not differ between regions (P > 0.99). Similarly, central versus peripheral activity after assay compound treatment showed no significant differences (P > 0.99, all time points). The intracorneal coefficient of variation for basal readings between two and four peripheral punches was 18.5% of the mean. Although peripheral samples displayed greater enzymatic activity than central samples (P < 0.05), similar to extracellular flux results, mitochondrial density did not differ between regions (P = 0.78). Extracellular flux analysis of oxygen and pH is a valid technique for characterizing metabolic activity of human corneal endothelium. This technique demonstrates high reproducibility, allows quantification of metabolic parameters using small quantities of live cells, and permits estimation of overall metabolic output. Neither oxygen consumption nor extracellular acidification differed between central and peripheral regions of transplant suitable corneas in this series. Our results show that endothelial cell health can be quantified biochemically in transplant suitable corneas.

Hypothermic preservation of corneas in a hyperkalaemic solution (CPTES): II. Extended storage in the presence of chondroitin sulphate.

PubMed Central

Taylor, M J; Hunt, C J; Madden, P W

1989-01-01

Periods of preservation for donor corneas, even for short times, are necessary to facilitate optimum conditions in penetrating keratoplasty. However, current techniques for corneal storage at low temperatures may not provide optimal conditions for maintaining tissue integrity. In particular, the ionic composition of the storage medium has received little attention since it has been assumed throughout that the normal complement of ions in tissue culture media will also be suitable for preservation at reduced temperatures. This study extends our previous investigations on the merits of using CPTES (corneal-potassium-TES), a potassium-rich balanced salt solution containing an impermeant anionic pH buffer (TES), as a storage solution specifically designed to prevent the loss of intracellular potassium and minimise endothelial cell swelling during the time that the normal regulatory processes are switched off. The effect of adding the natural polymer chondroitin sulphate (CS) as a colloid osmotic agent to the hyperkalaemic storage medium is now examined. Corneas stored in CPTES containing 2.5% chondroitin sulphate retained a very high level of structural and functional integrity after three, five, and seven days storage at 0 degrees C; furthermore, stromal swelling was restricted to only 21%. All corneas stored in CPTES + 2.5% CS showed active endothelial function by thinning efficiently at rates that were greater than those previously reported for rabbit corneas stored for similar lengths of time in either M-K medium or K-sol. The zwitterionic buffers TES and HEPES were interchangeable in the hyperkalaemic solution and were non-toxic to corneal endothelium at a concentration of 100 mM. These compounds offer excellent pH buffering in bicarbonate-free medium. Images PMID:2510816

Embryonic development of the cornea in the eye of the clearnose skate, Raja eglanteria: I. Stromal development in the absence of an endothelium

NASA Technical Reports Server (NTRS)

Conrad, G. W.; Paulsen, A. Q.; Luer, C. A.; Spooner, B. S. (Principal Investigator)

1994-01-01

Embryos of the clearnose skate, Raja eglanteria, develop in sea water at 20-22 degrees C, hatching after 82 +/- 4 days (Luer and Gilbert, Environ. Biol. Fishes, 13:161-171, 1985). Eyes develop as steadily enlarging spheres whose corneas have the same radius of curvature as the sclera. The cornea begins development as a 2-cell thick epithelium beneath which by Day 12 there is only a basal lamina and a wispy matrix separating it from the underlying lens. This matrix, modified by Day 16, is displaced on Day 22 by a few orthogonal plies of fibrillar primary stroma. Ply number increases to at least 13 by Day 30, reaching the final number of 20 +/- 2 by Day 42. Stromal fibroblasts (keratocytes) appear at the corneal periphery by Day 22, and in increased numbers by Day 30, a time at which no keratocytes are seen in the central stroma. However, by Day 40, many fibroblasts are present at the corneal periphery, invading the primary stroma between plies, occasionally reaching even the central cornea. By Day 53, keratocytes are present between all plies, from corneal periphery to center. Thickness of each ply in this secondary stroma increases, but the number of plies remains the same as in the primary stroma. Bowman's layer, non-invaded matrix beneath the epithelial basal lamina, is not evident until Day 53. Sutural fibers, first seen on Day 22, originate in the corneal epithelial basal lamina, traversing perpendicularly the plies of the primary stroma. Sutural fibers persist throughout development of the secondary stroma and into adulthood. In contrast to chicks, skate corneas remain transparent throughout development, and never form an endothelium.

Corneal neurotoxicity due to topical benzalkonium chloride.

PubMed

Sarkar, Joy; Chaudhary, Shweta; Namavari, Abed; Ozturk, Okan; Chang, Jin-Hong; Yco, Lisette; Sonawane, Snehal; Khanolkar, Vishakha; Hallak, Joelle; Jain, Sandeep

2012-04-06

The aim of this study was to determine and characterize the effect of topical application of benzalkonium chloride (BAK) on corneal nerves in vivo and in vitro. Thy1-YFP+ neurofluorescent mouse eyes were treated topically with vehicle or BAK (0.01% or 0.1%). Wide-field stereofluorescence microscopy was performed to sequentially image the treated corneas in vivo every week for 4 weeks, and changes in stromal nerve fiber density (NFD) and aqueous tear production were determined. Whole-mount immunofluorescence staining of corneas was performed with antibodies to axonopathy marker SMI-32. Western immunoblot analyses were performed on trigeminal ganglion and corneal lysates to determine abundance of proteins associated with neurotoxicity and regeneration. Compartmental culture of trigeminal ganglion neurons was performed in Campenot devices to determine whether BAK affects neurite outgrowth. BAK-treated corneas exhibited significantly reduced NFD and aqueous tear production, and increased inflammatory cell infiltration and fluorescein staining at 1 week (P < 0.05). These changes were most significant after 0.1% BAK treatment. The extent of inflammatory cell infiltration in the cornea showed a significant negative correlation with NFD. Sequential in vivo imaging of corneas showed two forms of BAK-induced neurotoxicity: reversible neurotoxicity characterized by axonopathy and recovery, and irreversible neurotoxicity characterized by nerve degeneration and regeneration. Increased abundance of beta III tubulin in corneal lysates confirmed regeneration. A dose-related significant reduction in neurites occurred after BAK addition to compartmental cultures of dissociated trigeminal ganglion cells. Although both BAK doses (0.0001% and 0.001%) reduced nerve fiber length, the reduction was significantly more with the higher dose (P < 0.001). Topical application of BAK to the eye causes corneal neurotoxicity, inflammation, and reduced aqueous tear production.

Corneal Neurotoxicity Due to Topical Benzalkonium Chloride

PubMed Central

Sarkar, Joy; Chaudhary, Shweta; Namavari, Abed; Ozturk, Okan; Chang, Jin-Hong; Yco, Lisette; Sonawane, Snehal; Khanolkar, Vishakha; Hallak, Joelle; Jain, Sandeep

2012-01-01

Purpose. The aim of this study was to determine and characterize the effect of topical application of benzalkonium chloride (BAK) on corneal nerves in vivo and in vitro. Methods. Thy1-YFP+ neurofluorescent mouse eyes were treated topically with vehicle or BAK (0.01% or 0.1%). Wide-field stereofluorescence microscopy was performed to sequentially image the treated corneas in vivo every week for 4 weeks, and changes in stromal nerve fiber density (NFD) and aqueous tear production were determined. Whole-mount immunofluorescence staining of corneas was performed with antibodies to axonopathy marker SMI-32. Western immunoblot analyses were performed on trigeminal ganglion and corneal lysates to determine abundance of proteins associated with neurotoxicity and regeneration. Compartmental culture of trigeminal ganglion neurons was performed in Campenot devices to determine whether BAK affects neurite outgrowth. Results. BAK-treated corneas exhibited significantly reduced NFD and aqueous tear production, and increased inflammatory cell infiltration and fluorescein staining at 1 week (P < 0.05). These changes were most significant after 0.1% BAK treatment. The extent of inflammatory cell infiltration in the cornea showed a significant negative correlation with NFD. Sequential in vivo imaging of corneas showed two forms of BAK-induced neurotoxicity: reversible neurotoxicity characterized by axonopathy and recovery, and irreversible neurotoxicity characterized by nerve degeneration and regeneration. Increased abundance of beta III tubulin in corneal lysates confirmed regeneration. A dose-related significant reduction in neurites occurred after BAK addition to compartmental cultures of dissociated trigeminal ganglion cells. Although both BAK doses (0.0001% and 0.001%) reduced nerve fiber length, the reduction was significantly more with the higher dose (P < 0.001). Conclusion. Topical application of BAK to the eye causes corneal neurotoxicity, inflammation, and reduced aqueous tear production. PMID:22410563

Potentiation of Femtosecond Laser Intratissue Refractive Index Shaping (IRIS) in the Living Cornea with Sodium Fluorescein

PubMed Central

Nagy, Lana J.; Ding, Li; Xu, Lisen; Knox, Wayne H.

2010-01-01

Purpose. To assess the effectiveness of intratissue refractive index shaping (IRIS) in living corneas and test the hypothesis that it can be enhanced by increasing the two-photon absorption (TPA) of the tissue. Methods. Three corneas were removed from adult cats and cut into six pieces, which were placed in preservative (Optisol-GS; Bausch & Lomb, Inc., Irvine, CA) containing 0%, 0.25%, 1%, 1.5%, or 2.5% sodium fluorescein (Na-Fl). An 800-nm Ti:Sapphire femtosecond laser with a 100-fs pulse duration and 80-MHz repetition rate was used to perform IRIS in each piece, creating several refractive index (RI) modification lines at different speeds (between 0.1 and 5 mm/s). The lines were 1 μm wide, 10 μm apart, and ∼150 μm below the tissue surface. The RI change of each grating was measured using calibrated, differential interference contrast microscopy. TUNEL staining was performed to assess whether IRIS or Na-Fl doping causes cell death. Results. Scanning at 0.1 mm/s changed the RI of undoped, living corneas by 0.005. In doped corneas, RI changes between 0.01 and 0.02 were reliably achieved with higher scanning speeds. The magnitude of RI changes attained was directly proportional to Na-Fl doping concentration and inversely proportional to the scanning speed used to create the gratings. Conclusions. IRIS can be efficiently performed in living corneal tissue. Increasing the TPA of the tissue with Na-Fl increased both the scanning speeds and the magnitude of RI changes in a dose-dependent manner. Ongoing studies are exploring the use of IRIS to alter the optical properties of corneal tissue in situ, over an extended period. PMID:19815735

A simple and non-contact optical imaging probe for evaluation of corneal diseases

NASA Astrophysics Data System (ADS)

Hong, Xun Jie Jeesmond; Shinoj, V. K.; Murukeshan, V. M.; Baskaran, M.; Aung, T.

2015-09-01

Non-contact imaging techniques are preferred in ophthalmology. Corneal disease is one of the leading causes of blindness worldwide, and a possible way of detection is by analyzing the shape and optical quality of the cornea. Here, a simple and cost-effective, non-contact optical probe system is proposed and illustrated. The probe possesses high spatial resolutions and is non-dependent on coupling medium, which are significant for a clinician and patient friendly investigation. These parameters are crucial, when considering an imaging system for the objective diagnosis and management of corneal diseases. The imaging of the cornea is performed on ex vivo porcine samples and subsequently on small laboratory animals, in vivo. The clinical significance of the proposed study is validated by performing imaging of the New Zealand white rabbit's cornea infected with Pseudomonas.

[Application value of corneal hysteresis in diagnosis and treatment of glaucoma].

PubMed

He, L Y; Liang, L; Zhu, M N

2017-02-11

Glaucoma is the first leading cause of irreversible blindness world widely, but the pathogenesis was still unclear. The collagen fibers from cornea and sclera connect to each other and both of them have similar extracellular matrix components. The biomechanical characteristics of optic nerve lamina cribrosa may associated with the biomechanical properties of the cornea. Therefore, the study of corneal physiological can indirectly reflex the compression and damage in optic nerve lamina cribrosa. The technical developments in corneal hysteresis examination had been updated these years constantly. Many researches implicated that low corneal hysteresis involved in pathogenesis and progression of glaucoma which refresh our recognition of the relationship between cornea and glaucoma. This review summarized the characteristics of corneal hysteresis, the examination and the connection with glaucoma to provide the reference for clinical work. (Chin J Ophthalmol, 2017, 53: 140-143) .

Measure the spatial distribution of corneal elasticity by combining femtosecond laser induced breakdown spectroscopy and acoustic radiation force elasticity microscope

NASA Astrophysics Data System (ADS)

Sun, Hui; Li, Xin; Hu, Mingyong

2017-08-01

The unique spatial distribution of corneal elasticity is shown by the nonhomogeneous structure of the cornea. It is critical to understanding how biomechanics control corneal stability and refraction and one way to do this job is non-invasive measurement of this distribution. Femtosecond laser pulses have the ability to induce optical breakdown and produced cavitation in the anterior and posterior cornea. A confocal ultrasonic transducer applied 6.5 ms acoustic radiation forcechirp bursts to the bubble at 1.5 MHz while monitoring bubble position using pulse-echoes at 20 MHz. The laser induced breakdown spectroscopy (LIBS) were measured in the anterior and posterior cornea with the plasmas that induced by the same femtosecond laser to see whether the laser induced plasmas signals will show relationship to Young's modulus.

A review of infrared laser energy absorption and subsequent healing in the cornea

NASA Astrophysics Data System (ADS)

Saunders, Latica L.; Johnson, Thomas E.; Neal, Thomas A.

2004-07-01

The purpose of this review is to compile information on the optical and healing properties of the cornea when exposed to infrared lasers. Our long-term goal is to optimize the treatment parameters for corneal injuries after exposure to infrared laser systems. The majority of the information currently available in the literature focuses on corneal healing after therapeutic vision correction surgery with LASIK or PRK. Only a limited amount of information is available on corneal healing after injury with an infrared laser system. In this review we will speculate on infrared photon energy absorption in corneal injury and healing to include the role of the tear layer. The aim of this review is to gain a better understanding of infrared energy absorption in the cornea and how it might impact healing.

Influence of changes in an eye's optical system on refraction

NASA Astrophysics Data System (ADS)

Bartkowska, Janina

1998-10-01

The optical system of eye is composed of cornea, lens, anterior chamber, and vitreous body. In the standard schematic eye there are 6 refracting surfaces. The changes of the curvature radii, of the distances between them, of the refractive indices influence the ametropia, refractive power of the eye and retinal image size. The influence of these changes can be appreciated by ray tracing or by an analytical method. There are presented simplified formulae for the differentials of ametropia and refractive power of the eye with respect to the surfaces curvatures, refracting power of cornea and lens, refractive indices. The relations are valid too for bigger changes if ametropia is measured in the cornea vertex. The formulae for the differentials with respect to distances, lens translation, eye axis length are valid if ametropia is measured in the object focus of the eye.

[Biological availability of ophthalmic drugs. 1. Increasing drug permeability in the cornea].

PubMed

Masteiková, R; Chalupová, Z; Savickiene, N

2004-03-01

Most eye diseases are treated by topical administration of ophthalmic preparations. Low ophthalmic bioavailability is due to a number of physiological and physicochemical factors. The main obstacle to the penetration of active ingredients to the eye is the layered character of the cornea. Formulation of ophthalmic preparations enabling easier penetration of this biological barrier thus ranks among the most effective ways of improving bioavailability. Penetrability of the cornea can be increased by the following methods: a) adjustment of the actual acidity in such a way that pH of the preparation makes it possible to produce the optimal portion of non-ionized particles; b) incorporation of absorption enhancers (non-ionic tensides, salts of bile acids, some antimicrobial substances, EDTA, cyclodextrins, etc.) into the composition of the preparation, and c) production of prodrugs or ionic pairs.

In vitro cell culture models to study the corneal drug absorption.

PubMed

Reichl, Stephan; Kölln, Christian; Hahne, Matthias; Verstraelen, Jessica

2011-05-01

Many diseases of the anterior eye segment are treated using topically applied ophthalmic drugs. For these drugs, the cornea is the main barrier to reaching the interior of the eye. In vitro studies regarding transcorneal drug absorption are commonly performed using excised corneas from experimental animals. Due to several disadvantages and limitations of these animal experiments, establishing corneal cell culture models has been attempted as an alternative. This review summarizes the development of in vitro models based on corneal cell cultures for permeation studies during the last 20 years, starting with simple epithelial models and moving toward complex organotypical 3D corneal equivalents. Current human 3D corneal cell culture models have the potential to replace excised animal corneas in drug absorption studies. However, for widespread use, the contemporary validation of existent systems is required.

Do heavy ions cause microlesions in cell membranes?

NASA Technical Reports Server (NTRS)

Koniarek, Jan P.; Worgul, Basil V.

1992-01-01

The microlesion question is investigated by monitoring the electrical potential difference across the endothelium of rat corneas in vitro before, during, and after irradiation. When the corneas were exposed to 1 Gy of Fe-56 ions (450 and 600 MeV/a.m.u.), no effect was detected on this parameter. These results suggest that direct physical damage to cell membranes, as predicted by the microlesion theory, does not take place.

Nanogel-based natural polymers as smart carriers for the controlled delivery of Timolol Maleate through the cornea for glaucoma.

PubMed

Ilka, Roya; Mohseni, Mojdeh; Kianirad, Mehran; Naseripour, Masood; Ashtari, Khadijeh; Mehravi, Bita

2018-04-01

Despite frequent scientific efforts, efficient ocular drug delivery is a major challenge for pharmaceutical scientists. Poor bioavailability of ophthalmic solutions can be overcome by using smart ophthalmic drug-delivery systems. In this research, loading and delivery of Timolol Maleate (TM) through the cornea by synthesized nanoparticles based on biopolymers (chitosan-alginate) were studied. The physico-chemical properties of these nanoparticles were characterized using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and dynamic light scattering (DLS). Loading and release were evaluated by a UV-vis spectrometer and the ex vivo permeation study was carried out using the Franz Diffusion Cell and fluorescent microscopy studies. The results indicated that morphology and size of nanoparticles were spherical and in the range of 80-100nm. The loading capacity and encapsulation efficiency were about 42% and 94% respectively. They illustrated a burst release in the first hour followed by a slower and more sustained drug release during the next 24h. Also, the results indicated that the cornea penetration of TM loaded in nanoparticles was twice than that of TM. Hence, this nanocarrier can be considered as a suitable candidate for controlled TM delivery and release through the cornea. Copyright © 2017 Elsevier B.V. All rights reserved.

Translational Immunoimaging and Neuroimaging Demonstrate Corneal Neuroimmune Crosstalk.

PubMed

Hamrah, Pedram; Seyed-Razavi, Yashar; Yamaguchi, Takefumi

2016-11-01

Corneal immunoimaging and neuroimaging approaches facilitate in vivo analyses of the cornea, including high-resolution imaging of corneal immune cells and nerves. This approach facilitates the analyses of underlying immune and nerve alterations not detected by clinical slit-lamp examination alone. In this review, we describe recent work performed in our translational ocular immunology center with a focus on "bench-to-bedside" and "bedside-to-bench" research. The ability to visualize dendritiform immune cells (DCs) in patients with laser in vivo confocal microscopy (IVCM), recently discovered in the central murine cornea, has allowed us to demonstrate their utility as a potential surrogate biomarker for inflammatory ocular surface diseases. This biomarker for inflammation allows the measurement of therapeutic efficacy of anti-inflammatory drugs and its utility as an endpoint in clinical trials with high interobserver agreement. IVCM image analyses from our studies has demonstrated a significant increase in DC density and size in ocular disease, a positive correlation between DC density and clinical signs and symptoms of disease and pro-inflammatory tear cytokines, and a strong negative correlation between DC density and subbasal nerve density. In conjunction with preclinical research investigating the inflammatory state in a partial or fully denervated cornea, our results indicated that corneal nerves are directly involved in the regulation of homeostasis and immune privilege in the cornea.

Modulating endogenous electric currents in human corneal wounds--a novel approach of bioelectric stimulation without electrodes.

PubMed

Reid, Brian; Graue-Hernandez, Enrique O; Mannis, Mark J; Zhao, Min

2011-03-01

To measure electric current in human corneal wounds and test the feasibility of pharmacologically enhancing the current to promote corneal wound healing. Using a noninvasive vibrating probe, corneal electric current was measured before and after wounding of the epithelium of donated postmortem human corneas. The effects of drug aminophylline and chloride-free solution on wound current were also tested. Unwounded cornea had small outward currents (0.07 μA/cm²). Wounding increased the current more than 5 fold (0.41 μA/cm²). Monitoring the wound current over time showed that it seemed to be actively regulated and maintained above normal unwounded levels for at least 6 hours. The time course was similar to that previously measured in rat cornea. Drug treatment or chloride-free solution more than doubled the size of wound currents. Electric current at human corneal wounds can be significantly increased with aminophylline or chloride-free solution. Because corneal wound current directly correlates with wound healing rate, our results suggest a role for chloride-free and/or aminophylline eyedrops to enhance healing of damaged cornea in patients with reduced wound healing such as the elderly or diabetic patient. This novel approach offers bioelectric stimulation without electrodes and can be readily tested in patients.

Stereo-PIV study of flow inside an eye under cataract surgery

NASA Astrophysics Data System (ADS)

Sakakibara, Jun; Yamashita, Masaki; Kobayashi, Tatsuya; Kaji, Yuichi; Oshika, Tetsuro

2012-04-01

We measured velocity distributions in the anterior chamber of porcine eyes under simulated cataract surgery using stereoscopic particle image velocimetry (stereo-PIV). The surface of the cornea was detected based on the images of laser-induced fluorescent light emitted from fluorescent dye solution introduced in a posterior chamber. A coaxial phacoemulsification procedure was simulated with standard size (standard coaxial phacoemulsification) and smaller (micro coaxial phacoemulsification) surgical instruments. In both cases, an asymmetric flow rate of irrigation was observed, although both irrigation ports had the same dimensions prior to insertion into the eye. In cases where the tip of the handpiece was placed farther away from the top of the cornea, i.e., closer to the crystalline lens, direct impingement of irrigation flow onto the cornea surface was avoided and the flow turned back toward the handpiece along the surface of the corneal endothelium. Viscous shear stress on the corneal endothelium was computed based on the measured mean velocity distribution. The maximum shear stress for most cases exceeded 0.1 Pa, which is comparable to the shear stress that caused detachment of the corneal endothelial cells reported by Kaji et al. in Cornea 24:S55-S58, (2005). When direct impingement of the irrigation flow was avoided, the shear stress was reduced considerably.

Role of Hemidivisional Corneal Topographic Astigmatisms (CorTs) in the Regularization and Reduction of Irregular Astigmatism.

PubMed

Alpins, Noel; Ong, James K Y; Stamatelatos, George

2018-03-01

To demonstrate how the concept of hemidivisional corneal topographic astigmatism (hemiCorT) enables the planning of hemidivisional corneal treatments to reduce irregularity and overall astigmatism. Whole-of-cornea corneal topographic astigmatism (CorT) is calculated from topography data derived from a corneal topographer or tomographer. The cornea is conceptually divided into 2 hemidivisions along the flat meridian of the CorT. For each hemidivision, hemiCorTs are calculated. The regularization treatment for each hemidivision is the treatment required to target the whole-of-cornea CorT, which is a symmetrical orthogonal corneal astigmatism. The regularization is then combined with astigmatism reduction treatment, which could be a conventional refractive treatment or a vector-planned treatment. For each hemidivision, the combined astigmatic effect of the regularization treatment and reduction treatment can be determined through double-angle vector summation. The 2 hemidivisional treatments together regularize and reduce corneal astigmatism. A theoretical pair of hemidivisional treatments is derived from an actual example of a cornea displaying idiopathic asymmetric nonorthogonal astigmatism. HemiCorTs allow for the design of hemidivisional corneal treatments of asymmetric nonorthogonal astigmatism. Such treatments should be suitable in the routine treatment of commonly occurring irregular astigmatism, while also allowing the spherical refractive error to be treated concurrently.

Effectiveness of Antimicrobial Peptide Immobilization for Preventing Perioperative Cornea Implant-Associated Bacterial Infection

PubMed Central

Tan, Xiao Wei; Goh, Tze Wei; Saraswathi, P.; Nyein, Chan Lwin; Setiawan, Melina; Riau, Andri; Lakshminarayanan, R.; Liu, Shouping; Tan, Donald; Beuerman, Roger W.

2014-01-01

Titanium (Ti) is a promising candidate biomaterial for an artificial corneal skirt. Antimicrobial peptide (AMP) immobilization may improve the bactericidal effect of the Ti substrate. In this study, we tested the bactericidal efficacy of a functionalized Ti surface in a rabbit keratitis model. A corneal stromal pocket was created by a femtosecond laser. The Ti films were then inserted into the pocket, and Staphylococcus aureus or Pseudomonas aeruginosa was inoculated into the pocket above the implant films. The corneas with Ti-AMP implants were compared with the corneas implanted with unprotected Ti by slit lamp observation and anterior segment optical coherence tomography (AS-OCT). Inflammatory responses were evaluated by bacterium counting, hematoxylin-eosin staining, and immunostaining. There was a lower incidence and a lesser extent of infection on rabbit corneas with Ti-AMP implants than on those with unprotected Ti implants. The bactericidal effect of AMP against S. aureus was comparable to that of postoperative prophylactic antibiotic treatment; hence, SESB2V AMP bound to the Ti implant provided functional activity in vivo, but its efficacy was greater against S. aureus than against P. aeruginosa. This work suggests that SESB2V AMP can be successfully functionalized in a rabbit keratitis model to prevent perioperative corneal infection. PMID:24957820

Combined use of a femtosecond laser and a microkeratome in obtaining thin grafts for Descemet stripping automated endothelial keratoplasty: an eye bank study.

PubMed

Murta, Joaquim N; Rosa, Andreia M; Quadrado, Maria Joao C; Russo, Ana D; Brito, Sergio S; Silva, Maria Fátima L

2013-01-01

To evaluate the use of a femtosecond laser combined with a microkeratome in the preparation of posterior corneal disks for Descemet stripping automated endothelial keratoplasty (DSAEK). This experimental study involved ultrathin DSAEK tissue preparation of 22 donor corneas unsuitable for transplantation. The first cut was performed with an Intralase® FS60 laser and the second cut with a Moria CBm 300-µm microkeratome. The thickness of the first cut was modified for each cornea to obtain a final graft thickness of less than 110 µm. Precut and postcut central pachymetry were performed with an ultrasonic pachymeter. Central endothelial cell density (ECD) was calculated before and 24 hours after tissue preparation. Final graft thickness was 105.0 ± 26.1 (SD) µm (range 65-117). The mean microkeratome head cut thickness was 324.5 ± 10.9 µm (range 310-345). Precut and postcut ECDs averaged 2250 ± 222 and 2093 ± 286 cells/mm2, respectively, representing 6.9% of cell loss. No corneas were perforated. Femtosecond FS60 lasers and Moria CBm 300-µm microkeratomes can be used sequentially to prepare consistently thin DSAEK grafts with no irregular cuts or cornea perforations.

T-style keratoprosthesis based on surface-modified poly (2-hydroxyethyl methacrylate) hydrogel for cornea repairs.

PubMed

Xiang, Jun; Sun, Jianguo; Hong, Jiaxu; Wang, Wentao; Wei, Anji; Le, Qihua; Xu, Jianjiang

2015-05-01

Corneal disease is a common cause of blindness, and keratoplasty is considered as an effective treatment method. However, there is a severe shortage of donor corneas worldwide. This paper presents a novel T-style design of a keratoprosthesis and its preparation methods, in which a mechanically and structurally effective artificial cornea is made based on a poly(2-hydroxyethyl methacrylate) hydrogel. The porous skirt was modified with hyaluronic acid and cationized gelatin, and the bottom of the optical column was coated with poly(ethylene glycol). The physical properties of the T-style Kpro were analyzed using ultraviolet and visible spectrophotometry and electron scanning microscopy. The surface chemical properties were characterized using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The surface modification in the spongy skirt promoted cell adhesion and produced a firm bond between the corneal tissue and the implant device, while the surface modification in the optic column resisted cell adhesion and prevented retroprosthetic membrane formation. Through improved surgical techniques, the novel T-style keratoprosthesis provides enough mechanical stability to facilitate long-term biointegration with the host environment. In vivo implantation experiments showed that the T-style keratoprosthesis is a promising cornea alternative for patients with severe limbal stem cell deficiency and corneal opacity. Copyright © 2015 Elsevier B.V. All rights reserved.

[Pathways of lanthanum ion transport across the posterior epithelium of the cornea in rabbits].

PubMed

Virnik, V L

1985-11-01

Morphological estimation of the barrier-transport properties of the posterior epithelium in the donor cornea has been performed in the most early time of fanoxic lesions of the cells. Ionized lanthanum, as an effective inhibitor of oxidative phosphorylation and simultaneously--as a marker of transport pathways for particles similar in their size to water molecule, is used in the investigation. The concentration gradient of rare-earth ions is produced by vital injection of lanthanum trichloride into the proper substance (stroma) of the cornea. From the microinjection focus the electron opaque marker is transported through the substrate of the basal substance of the stroma to the posterior epithelium and further along its intercellular and transcellular pathways into the anterior chamber of the eye. The rare-earth ions freely penetrate through the gap and tight junctions. Transcellular transport of lanthanum in the contents of the plasmolemmal microvesicles, vital absorption of the marker on the lateral and luminal parts of the plasmolemma and on the intracellular membranes, lesions of mitochondria and canaliculi of the endoplasmic reticulum of the anoxic character are stated. A suggestion is made on structural preservation of the pathways of convective and dissipative transition of the substance through the posterior epithelium of the cornea during the earliest time of the experimental anoxia.

Gene Therapy in the Cornea: 2005-present

PubMed Central

Mohan, Rajiv R.; Tovey, Jonathan C.K.; Sharma, Ajay; Tandon, Ashish

2011-01-01

Successful restoration of vision in human patients with gene therapy affirmed its promise to cure ocular diseases and disorders. The efficacy of gene therapy is contingent upon vector and mode of therapeutic DNA introduction into targeted cells/tissues. The cornea is an ideal tissue for gene therapy due to its ease of access and relative immune-privilege. Considerable progress has been made in the field of corneal gene therapy in last 5 years. Several new gene transfer vectors, techniques and approaches have evolved. Although corneal gene therapy is still in its early stages of development, the potential of gene-based interventions to treat corneal abnormalities have begun to surface. Identification of next generation viral and nanoparticle vectors, characterization of delivered gene levels, localization, and duration in the cornea, and significant success in controlling corneal disorders, particularly fibrosis and angiogenesis, in experimental animal disease models, with no major side effects have propelled gene therapy a step closer towards establishing gene-based therapies for corneal blindness. Recently, researchers have assessed the delivery of therapeutic genes for corneal diseases and disorders due to trauma, infections, chemical, mechanical, and surgical injury, and/or abnormal wound healing. This review provides an update on the developments in gene therapy for corneal diseases and discusses the barriers that hinder its utilization for delivering genes in the cornea. PMID:21967960

Corneal elastic anisotropy and hysteresis as a function of IOP assessed by optical coherence elastography

NASA Astrophysics Data System (ADS)

Li, Jiasong; Singh, Manmohan; Han, Zhaolong; Wu, Chen; Raghunathan, Raksha; Liu, Chih-Hao; Nair, Achuth; Noorani, Shezaan; Aglyamov, Salavat R.; Twa, Michael D.; Larin, Kirill V.

2016-03-01

The mechanical anisotropic properties of the cornea can be an important indicator for determining the onset and severity of different diseases and can be used to assess the efficacy of various therapeutic interventions, such as cross-linking and LASIK surgery. In this work, we introduce a noncontact method of assessing corneal mechanical anisotropy as a function of intraocular pressure (IOP) using optical coherence elastography (OCE). A focused air-pulse induced low amplitude (<10 μm) elastic waves in fresh porcine corneas in the whole eye-globe configuration in situ. A phase-stabilized swept source optical coherence elastography (PhS-SSOCE) system imaged the elastic wave propagation at stepped radial angles, and the OCE measurements were repeated as the IOP was cycled. The elastic wave velocity was then quantified to determine the mechanical anisotropy and hysteresis of the cornea. The results show that the elastic anisotropy at the corneal of the apex of the cornea becomes more pronounced at higher IOPs, and that there are distinct radial angles of higher and lower stiffness. Due to the noncontact nature and small amplitude of the elastic wave, this method may be useful for characterizing the elastic anisotropy of ocular and other tissues in vivo completely noninvasively.

Shortage in the face of plenty: improving the allocation of corneas for transplantation.

PubMed

de By, T M M H

2003-01-01

As there are plenty of potential corneal donors, theoretically there should be no shortage of corneal grafts. Practically, however, shortages have been a problem, especially with HLA-matched corneas, which are increasingly requested. This increase requires evaluation and proper adaptation to meet all requests in the future. BIS' allocation data for corneal transplants were analyzed for the years 1998 through 2001. Allocation of matched corneas almost doubled in 2000. The waiting list for matched grafts could be reduced by one third. A steep increase in demand for matched grafts could be noted by the year 2000. This signifies that the results of positive matching studies have ultimately been taken notice of by the ophthalmic surgeons with consequent adaptation of their graft orders. BIS has mostly been able to cope with these rapidly changing and increasing demands, but more efforts are continuously needed to supply those typed and matched corneas within short time. Our next step will be to offer HLA-DR matching routinely in addition to the current HLA-A/-B matching. We are looking forward with great interest to the introduction of even more specified matching algorithms in the not too far distant future, and are prepared to meet all the requirements which might be associated with such challenges.

Three-dimensional and topographic relationships between the orbital margins with reference to assessment of eyeball protrusion

PubMed Central

Shin, Kang-Jae; Lee, Shin-Hyo; Koh, Ki-Seok

2017-01-01

This study investigated the topographic relationships among the eyeball and four orbital margins with the aim of identifying the correlation between orbital geometry and eyeball protrusion in Koreans. Three-dimensional (3D) volume rendering of the face was performed using serial computed-tomography images of 141 Koreans, and several landmarks on the bony orbit and the cornea were directly marked on the 3D volumes. The anterior-posterior distances from the apex of the cornea to each orbital margin and between the orbital margins were measured in both eyes. The distances from the apex of the cornea to the superior, medial, inferior, and lateral orbital margins were 5.8, 5.8, 12.0, and 17.9 mm, respectively. Differences between sides were observed in all of the orbital margins, and the distances from the apex of the cornea to the superior and inferior orbital margins were significantly greater in females than in males. The anterior-posterior distance between the superior and inferior orbital margins did not differ significantly between males (6.3 mm) and females (6.2 mm). The data obtained in this study will be useful when developing practical guidelines applicable to forensic facial reconstruction and orbitofacial surgeries. PMID:28417054

Rebamipide increases mucin-like substance contents and periodic acid Schiff reagent-positive cells density in normal rabbits.

PubMed

Urashima, Hiroki; Takeji, Yasuhiro; Okamoto, Takashi; Fujisawa, Shigeki; Shinohara, Hisashi

2012-06-01

The effects of rebamipide on the number of periodic acid Schiff reagent (PAS)-positive cells in the conjunctiva, the mucin content in the cornea and conjunctiva of normal rabbits, and desiccation-induced corneal damage in vivo were examined. Rebamipide (0.1%-3%) was applied 6 times a day for 14 days, and the PAS-positive cell count in the bulbar conjunctiva was measured by impression cytology. The amount of conjunctival and corneal mucin-like substances was measured by Alcian blue binding. The corneal damage model was created by desiccation from air flow at room temperature. The level of corneal damage was determined by scoring the area stained with rose bengal and fluorescein dye. Rebamipide increased the number of PAS-positive cells in the conjunctiva when instilled at concentrations of 0.3% or higher, and 1% rebamipide increased the amount of mucin-like substances of the conjunctiva and cornea. Moreover, 1% rebamipide was also found to lower the rose bengal scores of the cornea in the corneal damage model by desiccation. Rebamipide is a possible candidate drug for treatment of cornea and conjunctival epithelial damage due to its mucin-like substance increasing action, for instance, in the treatment of dry eye disease.

[Intraoperative Optical Coherence Tomography (MI-OCT) for the Treatment of Corneal Dystrophies].

PubMed

Siebelmann, Sebastian; Matthaei, Mario; Heindl, Ludwig M; Bachmann, Björn O; Cursiefen, Claus

2018-06-01

The surgical treatment of corneal dystrophies develops rapidly as the use of lamellar corneal grafting techniques continue. While penetrating keratoplasty was the gold standard for treating a variety of dystrophies a few years ago, the affected layers of the cornea can, nowadays, be selectively replaced or ablated using laser technology. Of particular importance for these methods is optical coherence tomography, which has recently been integrated into surgical microscopes (MI-OCT). Literature overview from PubMed and Google.scholar.de supplemented with own imaging data. The MI-OCT enables the intraoperative real-time monitoring of different ophthalmic surgical procedures, such as deep anterior lamellar keratoplasty, Descemet's membrane endothelial keratoplasty, as well as minimally-invasive procedures, such as phototherapeutic keratectomy. In addition, it enables an evaluation of the cornea, but also of structures of the anterior chamber, in situations in which the cornea, for example, is clouded by an edema. Microscope-integrated, intraoperative optical coherence tomography (MI-OCT) represents a useful supplement to the normal surgical microscope. It is superior to the sole surgical microscope, especially in already severely clouded corneas, and represents a sensible supplement, especially for novel lamellar transplantation procedures. Prospective randomized trials are necessary to increase safety and efficacy when using MI-OCT for different indications. Georg Thieme Verlag KG Stuttgart · New York.

Study of transmittance and reflectance spectra of the cornea and the sclera in the THz frequency range

NASA Astrophysics Data System (ADS)

Iomdina, Elena N.; Goltsman, Gregory N.; Seliverstov, Sergey V.; Sianosyan, Alisa A.; Teplyakova, Kseniya O.; Rusova, Anastasia A.

2016-09-01

An adequate water balance (hydration extent) is one of the basic factors of normal eye function, including its external shells: the cornea and the sclera. Adequate control of corneal and scleral hydration is very important for early diagnosis of a variety of eye diseases, stating indications for and contraindications against keratorefractive surgeries and the choice of contact lens correction solutions. THz systems of creating images in reflected beams are likely to become ideal instruments of noninvasive control of corneal and scleral hydration degrees. This paper reports on the results of a study involving transmittance and reflectance spectra for the cornea and the sclera of rabbit and human eyes, as well as those of the rabbit eye, in the frequency range of 0.13 to 0.32 THz. The dependence of the reflectance coefficient of these tissues on water mass percentage content was determined. The experiments were performed on three corneas, three rabbit scleras, two rabbit eyes, and three human scleras. The preliminary results demonstrate that the proposed technique, based on the use of a continuous THz radiation, may be utilized to create a device for noninvasive control of corneal and scleral hydration, which has clear potential of broad practical application.

Assessment of corneal hydration sensing in the terahertz band: in vivo results at 100 GHz

NASA Astrophysics Data System (ADS)

Bennett, David; Taylor, Zachary; Tewari, Pria; Sung, Sijun; Maccabi, Ashkan; Singh, Rahul; Culjat, Martin; Grundfest, Warren; Hubschman, Jean-Pierre; Brown, Elliott

2012-09-01

Terahertz corneal hydration sensing has shown promise in ophthalmology applications and was recently shown to be capable of detecting water concentration changes of about two parts in a thousand in ex vivo corneal tissues. This technology may be effective in patient monitoring during refractive surgery and for early diagnosis and treatment monitoring in diseases of the cornea. In this work, Fuchs dystrophy, cornea transplant rejection, and keratoconus are discussed, and a hydration sensitivity of about one part in a hundred is predicted to be needed to successfully distinguish between diseased and healthy tissues in these applications. Stratified models of corneal tissue reflectivity are developed and validated using ex vivo spectroscopy of harvested porcine corneas that are hydrated using polyethylene glycol solutions. Simulation of the cornea's depth-dependent hydration profile, from 0.01 to 100 THz, identifies a peak in intrinsic reflectivity contrast for sensing at 100 GHz. A 100 GHz hydration sensing system is evaluated alongside the current standard ultrasound pachymetry technique to measure corneal hydration in vivo in four rabbits. A hydration sensitivity, of three parts per thousand or better, was measured in all four rabbits under study. This work presents the first in vivo demonstration of remote corneal hydration sensing.

Transdifferentiation from cornea to lens in Xenopus laevis depends on BMP signalling and involves upregulation of Wnt signalling

PubMed Central

2011-01-01

Background Surgical removal of the lens from larval Xenopus laevis results in a rapid transdifferention of central corneal cells to form a new lens. The trigger for this process is understood to be an induction event arising from the unprecedented exposure of the cornea to the vitreous humour that occurs following lens removal. The molecular identity of this trigger is unknown. Results Here, we have used a functional transgenic approach to show that BMP signalling is required for lens regeneration and a microarray approach to identify genes that are upregulated specifically during this process. Analysis of the array data strongly implicates Wnt signalling and the Pitx family of transcription factors in the process of cornea to lens transdifferentiation. Our analysis also captured several genes associated with congenital cataract in humans. Pluripotency genes, in contrast, were not upregulated, supporting the idea that corneal cells transdifferentiate without returning to a stem cell state. Several genes from the array were expressed in the forming lens during embryogenesis. One of these, Nipsnap1, is a known direct target of BMP signalling. Conclusions Our results strongly implicate the developmental Wnt and BMP signalling pathways in the process of cornea to lens transdifferentiation (CLT) in Xenopus, and suggest direct transdifferentiation between these two anterior eye tissues. PMID:21896182

Recipient Risk Factors for Graft Failure in the Cornea Donor Study

PubMed Central

Sugar, Alan; Tanner, Jean Paul; Dontchev, Mariya; Tennant, Brad; Schultze, Robert L.; Dunn, Steven P.; Lindquist, Thomas D.; Gal, Robin L.; Beck, Roy W.; Kollman, Craig; Mannis, Mark J.; Holland, Edward J.

2009-01-01

Purpose Identify recipient factors which may be related to risk of corneal graft failure Design Multi-center prospective, double-masked, controlled clinical trial Participants 1090 subjects undergoing corneal transplantation for a moderate risk condition (principally Fuchs’ dystrophy or pseudophakic corneal edema) Methods Donor corneas were assigned using a random approach without respect to recipient factors, and surgeons were masked to information about the donor cornea including donor age. Surgery and post-operative care were performed according to the surgeons’ usual routines and subjects were followed for five years. Baseline factors were evaluated for their association with graft failure. Main Outcome Measures Graft failure, defined as a regraft or a cloudy cornea that was sufficiently opaque to compromise vision for a minimum of three consecutive months. Results Preoperative diagnosis of pseudophakic/aphakic corneal edema increased graft failure risk approximately 4-fold compared with Fuchs’ dystrophy (27% vs. 7%). Prior glaucoma surgery with preoperative glaucoma medication use substantially increased the graft failure rate. Factors not strongly associated with graft failure included age, gender, diabetes, smoking, and graft size. Conclusion The risk of graft failure is significantly increased in eyes with pseudophakic or aphakic corneal edema compared with Fuchs’ dystrophy, independent of lens status, and in eyes with a history of glaucoma. PMID:19395036

Toxicological effects and recovery of the corneal epithelium in Cyprinus carpio communis Linn. exposed to monocrotophos: an scanning electron microscope study.

PubMed

Uppal, Ravneet Kaur; Johal, Mohinder Singh; Sharma, Madan Lal

2015-05-01

This study was conducted based on the evidence of fish habitats in North India being affected by organophosphate pesticides draining from agricultural fields into bodies of water, especially during the rainy season. Various tissues of fish such as scales, gills ovaries, kidney, and liver have been studied from the toxicological point of view, but the toxicological effects of aquatic pollutants on fish cornea have not been investigated to date. We conducted comparative toxicological studies on the cornea of Cyprinus carpio communis using two sublethal (0.038 and 0.126 ppm) concentrations of monocrotophos pesticide for 30 days. Corneas from all the groups were evaluated by a scanning electron microscope. The fish exposed to the monocrotophos pesticide developed corneal necrosis due to the formation of crystalloid-like structures, thinning and shrinkage of microridges on the corneal epithelium. After 30 days, fish from the monocrotophos-treated tank were transferred to normal environmental conditions. After 60 days under natural condition, epithelial cells did not fully recover. In conclusion, exposure to monocrotophos induces irreversible changes in the cornea of C. carpio communis. As fish and mammalian visual systems share many similarities, the reported finding may offer useful insights for further toxicological and ophthalmological studies in humans. © 2013 American College of Veterinary Ophthalmologists.

Damage Threshold of In Vivo Rabbit Cornea by 2 micron Laser Irradiation

DTIC Science & Technology

2007-01-01

in laser injury experiments? Implications for human exposure limits. Health Phys 2002; 82(3):335-347. 11. Siegman AE, Sasnett MW, Johnston TF. Choice... Laser Irradiation DISTRIBUTION: Approved for public release, distribution unlimited This paper is part of the following report: TITLE: Conference on...part numbers comprise the compilation report: ADP023676 thru ADP023710 UNCLASSIFIED Damage Threshold of In Vivo Rabbit Cornea by 2 gm Laser Irradiation

An Operational Evaluation of Extended-Wear Soft Contact Lenses in an Armored Division.

DTIC Science & Technology

1987-08-01

has been demonstrated in the tarsal portion of the palpebral conjunctiva , usually of the upper . lid (Korb et al., 1983). Termed giant papillary...segment of the eye; any disease that affected the cornea, conjunctiva or sclera; corneal hypoesthesia; low tear breakup time or insufficient...evaluation of the following: lids, conjunctiva , limbus, cornea, anterior chamber, and tarsal plate. . 14 v\\Md rn, , !,F~l *O NO .. W,% Quantification of

Entertainment-education and recruitment of cornea donors: the role of emotion and issue involvement.

PubMed

Bae, Hyuhn-Suhck

2008-01-01

This study examined the role of emotional responses and viewer's level of issue involvement to an entertainment-education show about cornea donation in order to predict intention to register as cornea donors. Results confirmed that sympathy and empathy responses operated as a catalyst for issue involvement, which emerged as an important intermediary in the persuasion process. Issue involvement also was found to be a common causal antecedent of attitude, subjective norm, and perceived behavioral control, the last two of which predict intentions unlike attitude, which does not. The revised path model confirmed that involvement directly influences intention. The findings of this study suggest that adding emotion and involvement in the Theory of Planned Behavior (TPB) enhances the explanatory power of the theory in predicting intentions, which indicates the possibility of combining the Elaboration Likelihood Model (ELM) and the TPB in the prediction of human behaviors.

THz Medical Imaging: in vivo Hydration Sensing

PubMed Central

Taylor, Zachary D.; Singh, Rahul S.; Bennett, David B.; Tewari, Priyamvada; Kealey, Colin P.; Bajwa, Neha; Culjat, Martin O.; Stojadinovic, Alexander; Lee, Hua; Hubschman, Jean-Pierre; Brown, Elliott R.; Grundfest, Warren S.

2015-01-01

The application of THz to medical imaging is experiencing a surge in both interest and federal funding. A brief overview of the field is provided along with promising and emerging applications and ongoing research. THz imaging phenomenology is discussed and tradeoffs are identified. A THz medical imaging system, operating at ~525 GHz center frequency with ~125 GHz of response normalized bandwidth is introduced and details regarding principles of operation are provided. Two promising medical applications of THz imaging are presented: skin burns and cornea. For burns, images of second degree, partial thickness burns were obtained in rat models in vivo over an 8 hour period. These images clearly show the formation and progression of edema in and around the burn wound area. For cornea, experimental data measuring the hydration of ex vivo porcine cornea under drying is presented demonstrating utility in ophthalmologic applications. PMID:26085958

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kitai, M S; Semchishen, A V; Semchishen, V A

The optical quality of the eye cornea surface after performing the laser vision correction essentially depends on the characteristic roughness scale (CRS) of the ablated surface, which is mainly determined by the absorption coefficient of the cornea at the laser wavelength. Thus, in the case of using an excimer ArF laser (λ = 193 nm) the absorption coefficient is equal to 39000 cm{sup -1}, the darkening by the dissociation products takes place, and the depth of the roughness relief can be as large as 0.23 mm. Under irradiation with the Er : YAG laser (λ = 2940 nm) the clearingmore » is observed due to the rupture of hydrogen bonds in water, and the relief depth exceeds 1 μm. It is shown that the process of reepithelization that occurs after performing the laser vision correction leads to the improvement of the optical quality of the cornea surface. (interaction of laser radiation with matter)« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Skrinnik, A.V.

The author attempts to obtain direct proof of increased permeability of the cornea and enhanced sorption properties of the tissue structures and refractive media of the eye under the influence of a pulsed magnetic field (MF). In the course of the investigation, the method of radioactive indication of two substances (/sup 35/S-streptomycin and /sup 75/Se-methionine), widely used in opthalmologic practice, was used. The radioactivity of the working solution of the preparations was 300,000 cpm in 0.1 ml. Changes in the permeability of the cornea were assessed on the basis of changes in radioactivity of the aqueous humor. The concentration ofmore » /sup 35/S-streptomycin in aqueous humor of the animals immediately after exposure to MF was significantly higher than in the control. Analysis of permeability of the cornea for /sup 75/Se-methionine also showed increased penetration of radioactivity into the aqueous. The results are evidence of the greater penetrating power of /sup 75/Se-methionine.« less

Novel monitoring of corneal surface hydration during photorefractive keratectomy using pulsed photothermal radiometry: in-vitro study

NASA Astrophysics Data System (ADS)

Kawauchi, Satoko; Matsuyama, Hiroko; Obara, Minoru; Ishihara, Miya; Arai, Tsunenori; Kikuchi, Makoto; Katoh, Masayoshi

1997-05-01

We developed novel monitoring methodology for corneal surface hydration during photorefractive keratectomy (PRK) in order to solve undercorrection issue at the central part of cornea (Central island). We employed pulsed photothermal radiometry to monitor corneal surface hydration. We performed two experiments; gelatin gel experiments and porcine cornea experiments in vitro. In the case of the gelatin gel experiments, the e-folding decay time of transient infrared radiation waveform from the ArF laser irradiated surface was prolonged from 420 microsecond(s) to 30 ms with decreasing gelatin density from 15% to 0.15%. These measured e-folding decay times were good agreements with theoretical calculations. Using porcine cornea, we observed the e-folding decay time increase during the series of ArF excimer laser irradiations. Our method may be available to know ablation efficiency change to improve the controllability of refractive correction on the PRK.

Assessment of the Dutch organ-culture system of corneal preservation within the Eye Bank of South Australia.

PubMed

Williams, K A; Noack, L M; Alfrich, S J; Danz, R; Erickson, S A; Coster, D J

1988-02-01

Thirteen per cent of all corneas harvested by the Eye Bank of South Australia during 1986 were discarded because storage time in McCarey-Kaufman medium exceeded four days. We have therefore examined the suitability of the Dutch method of long-term corneal storage for our purposes. Twenty-two human corneas that had been discarded from the Eye Bank were assessed using the trypan blue-sucrose staining technique, and then placed into long-term storage for 15 to 17 days. They were then reassessed by vital dye staining before permanent flat-mounts were prepared for silver staining of the endothelium. A good correlation (albeit subjective) was found between the non-destructive and destructive techniques of endothelial cell assessment. Those corneas that failed to survive organ culture storage were easily detected. The Dutch system of corneal preservation and post-storage assessment seems well-suited to Australian eye-banking.

Social influence of a religious hero: the late Cardinal Stephen Kim Sou-hwan's effect on cornea donation and volunteerism.

PubMed

Bae, Hyuhn-Suhck; Brown, William J; Kang, Seok

2011-01-01

This study examined the mediated influence of a celebrated religious hero in South Korea, Cardinal Stephen Kim, through two forms of involvement--parasocial interaction and identification--on intention toward cornea donation and volunteerism, and it investigated how the news media diffused of his death. A structural equation modeling analysis with a Web-based voluntary survey of more than 1,200 people in South Korea revealed a multistep social influence process, beginning with parasocial interaction with Cardinal Kim, leading to identification with him, which predicted intention toward cornea donation and volunteerism. Additional investigations found that news of Cardinal Kim's death diffused rapidly through media and interpersonal communication. Results of this study demonstrate that religious leaders who achieve a celebrity hero status can prompt public discussion of important issues rather quickly through extensive media coverage, enabling them to promote prosocial behavior and positively affect public health.

Co:MgF2 laser ablation of tissue: effect of wavelength on ablation threshold and thermal damage.

PubMed

Schomacker, K T; Domankevitz, Y; Flotte, T J; Deutsch, T F

1991-01-01

The wavelength dependence of the ablation threshold of a variety of tissues has been studied by using a tunable pulsed Co:MgF2 laser to determine how closely it tracks the optical absorption length of water. The Co:MgF2 laser was tuned between 1.81 and 2.14 microns, a wavelength region in which the absorption length varies by a decade. For soft tissues the ablation threshold tracks the optical absorption length; for bone there is little wavelength dependence, consistent with the low water content of bone. Thermal damage vs. wavelength was also studied for cornea and bone. Thermal damage to cornea has a weak wavelength dependence, while that to bone shows little wavelength dependence. Framing-camera pictures of the ablation of both cornea and liver show explosive removal of material, but differ as to the nature of the explosion.

Human Asymptomatic Epitope Peptide/CXCL10-Based Prime/Pull Vaccine Induces Herpes Simplex Virus-Specific Gamma Interferon-Positive CD107+ CD8+ T Cells That Infiltrate the Cornea and Trigeminal Ganglia of Humanized HLA Transgenic Rabbits and Protect against Ocular Herpes Challenge.

PubMed

Khan, Arif A; Srivastava, Ruchi; Vahed, Hawa; Roy, Soumyabrata; Walia, Sager S; Kim, Grace J; Fouladi, Mona A; Yamada, Taikun; Ly, Vincent T; Lam, Cynthia; Lou, Anthony; Nguyen, Vivianna; Boldbaatar, Undariya; Geertsema, Roger; Fraser, Nigel W; BenMohamed, Lbachir

2018-06-13

Herpes simplex virus 1 (HSV-1) is a prevalent human pathogen that infects the cornea causing potentially blinding herpetic disease. A clinical herpes vaccine is still lacking. In the present study, a novel prime/pull vaccine was tested in Human Leukocyte Antigen- (HLA-) transgenic rabbit model of ocular herpes (HLA Tg rabbit). Three asymptomatic (ASYMP) peptide epitopes were selected from the HSV-1 membrane glycoprotein C (UL44 400-408 ), the DNA replication binding helicase (UL9 196-204 ), and the tegument protein (UL25 572-580 ), all preferentially recognized by CD8 + T cells from "naturally protected" HSV-1-seropositive healthy ASYMP individuals (who never had recurrent corneal herpetic disease). HLA Tg rabbits were immunized with a mixture of these three ASYMP CD8 + T cell peptide epitopes (UL44 400-408 , UL9 196-204 and UL25 572-580 ), delivered subcutaneously with CpG 2007 adjuvant (prime). Fifteen days later, half of the rabbits received a topical ocular treatment with a recombinant neurotropic AAV8 vector, expressing the T cell-attracting CXCL10 chemokine (pull). The frequency, function of HSV-specific CD8 + T cells induced by the prime/pull vaccine were assessed in peripheral blood, cornea, and trigeminal ganglia (TG). Compared to peptides alone, the peptides/CXCL10 prime/pull vaccine generated frequent polyfunctional gamma interferon-positive (IFN-γ + ) CD107 + CD8 + T cells that infiltrated both the cornea and TG. CD8 + T cells mobilization into cornea and TG of prime/pull- vaccinated rabbits was associated with a significant reduction in corneal herpes infection and disease following an ocular HSV-1 challenge (McKrae). These findings draw attention to the novel prime/pull vaccine strategy to mobilize anti-viral CD8 + T cells into tissues protecting them against herpes infection and disease. IMPORTANCE There is an urgent need for a vaccine against widespread herpes simplex virus infections. The present study demonstrates that immunization of HLA transgenic rabbits with a peptide/CXCL10 prime/pull vaccine triggered mobilization of HSV-specific CD8 + T cells locally in the cornea and TG, the sites of acute and latent herpes infections. Mobilization of antiviral CD8 + T cells into cornea and TG of rabbits that received the prime/pull vaccine was associated with protection against ocular herpes infection and disease following an ocular HSV-1 challenge. These results highlight the importance of the prime/pull vaccine strategy to bolster the number and function of protective CD8 + T cells within infected tissues. Copyright © 2018 American Society for Microbiology.

[Morphological characteristics of cornea in patients with vernal keratoconjunctivitis by in vivo laser scanning confocal microscopy].

PubMed

Le, Qi-hua; Hong, Jia-xu; Zhu, Wen-qing; Sun, Xing-huai; Xu, Jian-jiang

2011-05-01

To explore the morphological characteristics on cornea in patients with vernal keratoconjunctivitis (VKC) by the application of in vivo laser scanning confocal microscopy (LSCM). The experimental design was retrospective observation case series (case control study). Twenty-six patients, each diagnosed as bilateral VKC, were enrolled in the study, among which 13 were tarsal form, 5 were bulbar form and the rest were mixed form. Nine patients had the clinical course less than one year, eight subjects longer than three years, and the rest between them. Another twenty-six healthy volunteers with matching age and gender were selected as normal control. All participants had their right eyes examined with the in vivo confocal microscopy (HRT II/RCM). Central cornea and superior peripheral cornea were chosen as the examination points. The images were recorded automatically and cellular density of each layer was analyzed by installed software. Software ImageJ was utilized to analyze the density, diameter, branch number and tortuosity of subbasal nerve fiber in VKC patients. Independent t test was performed to assess the differences on cellular density between VKC patients and normal control, as well as those between central and peripheral cornea in VKC patients. Fisher chi-square test was used to compare the infiltration rate of Langerhans cells in corneal epithelium between VKC patients and controls. ANOVA was applied to assess the differences in cellular density among three subtypes, as well as among different duration of VKC. Independent t-test and chi-square test were applied to analyze the parameters of subbasal nerve fiber. The morphological changes in cornea included the absence of superficial hyperreflective polygonal epithelial cells, infiltration of Langerhans cells in and(or) underneath corneal epithelium and activation of keratocytes in anterior stroma. Corneal epithelium conjunctivalization and stromal neovascularization could be identified in patients with corneal neovascular epithelium. Longitudinal or oblique dark striae could be found in the posterior stroma in patients with complicated keratoconus. The density of epithelial cells at central and peripheral cornea in healthy controls were (6033.1 ± 998.7) cells/mm(2) and (6098.4 ± 298.3) cells/mm(2), while that in VKC patients were (5972.2 ± 1148.2) cells/mm(2) and (6178.5 ± 318.9) cells/mm(2) respectively, the differences being no statistical significant between them (t = 1.191, 1.011; P = 0.238, 0.318). However, it's found in VKC patients that cellular density at peripheral cornea was significantly higher than that at central area (t = 2.249, P = 0.03). The density of anterior stromal cells at central and peripheral cornea in healthy controls was (1001.4 ± 125.3) cells/mm(2) and (924.6 ± 201.4) cells/mm(2), while that in VKC patients was (1184.5 ± 115.3) cells/mm(2) and (1101.4 ± 151.1) cells/mm(2), the difference bearing no statistical significance (t = 6.617, 3.439; P = 0.001). The density of posterior stromal cells in normal subjects and VKC patients was (537.7 ± 42.6) cells/mm(2) and (548.7 ± 79.8) cells/mm(2), that of endothelial cells was (2985.7 ± 401.2) cells/mm(2) and (3021.5 ± 383.3) cells/mm(2), respectively, neither difference had statistical significance (t = 0.174, 1.112; P = 0.864, 0.282). Langerhans cell infiltration could be identified in 61.5% (16 cases) VKC patients, which was significantly higher than normal control (2 cases, 7.7%) (χ(2) = 12.49, P = 0.001). Furthermore, much intense Langerhans cells infiltration was found in bulbar form and mix form than tarsal form. (t = 6.617, P = 0.001). The density and diameter of subbasal nerve fiber in VKC patients decreased significantly than those in normal subjects, whereas the tortuosity increased significantly. The morphological changes of cornea in VKC patients mainly involve corneal epithelium, subbasal nerve fiber and anterior stroma. In vivo LSCM is helpful in discriminating the subtypes of VKC.

Association of fluorescein anterior corneal mosaic and corneal K-structures by in vivo laser confocal microscopy in patients with keratoconus.

PubMed

Kobayashi, Akira; Yokogawa, Hideaki; Mori, Natsuko; Masaki, Toshinori; Sugiyama, Kazuhisa

2017-01-01

To report the in vivo laser confocal microscopy findings of corneas with keratoconus, with special attention to abnormality of Bowman's layer and sub-Bowman's fibrous structures (Kobayashi-structures [K-structures]). Sixteen keratoconic eyes in 8 consecutive patients with keratoconus (4 males, 4 females, mean age, 41.1 years) were included in this study. Slit-lamp biomicroscopic photos were taken with or without fluorescein staining. The existence of anterior corneal mosaic (ACM) after eyelid rubbing under fluorescein staining was documented. In vivo laser confocal microscopic examinations were performed for all patients in both the central cone and the peripheral cornea to examine the existence of K-structures. According to the Amsler-Krumeich scale, the eyes were graded as follows: stage 1 (n=3), stage 2 (n=1), stage 3 (n=1), and stage 4 (n=11). ACM was observed in 7 eyes (61.1%) in the cone area and 16 eyes (100%) in the peripheral cornea among all keratoconic eyes enrolled in this study. In addition, K-structures were observed in the 7 eyes (61.1%) and 16 eyes (100%) in the peripheral cornea among all keratoconic eyes. The presence of the K-structures was completely matched (100%) with the presence of ACM in both the central cone and the peripheral cornea. In 11 eyes with stage 4 keratoconus, ACM and K-structure was absent in 9 eyes (81.8%) in the cone area. On the contrary, in 5 eyes with mild-to-moderate keratoconus (grade 1 to 3), ACM and K-structure was present in all eyes (100%) in the cone area. The absent ratio of ACM and K-structures in the cone area was significantly higher in stage 4 severe keratoconus compared to mild-to-moderate keratoconus (grade 1 to 3) (Fisher, P =0.005). The existence of ACM and K-structures in both the central cone and the peripheral cornea showed perfect accord in patients with keratoconus, indicating a strong association of ACM and K-structures in patients with keratoconus. With the progress of the keratoconus, it seemed that ACM and K-structure progressively disappeared, suggesting Bowman's layer abnormalities due to keratoconus. Further study in larger groups of patients with keratoconus is required to fully understand the significance of ACM/K-structures in keratoconic eyes and their association with Bowman's layer.

Hyaluronic acid coated poly-epsilon-caprolactone nanospheres deliver high concentrations of cyclosporine A into the cornea.

PubMed

Yenice, Irem; Mocan, Mehmet C; Palaska, Erhan; Bochot, Amélie; Bilensoy, Erem; Vural, Imran; Irkeç, Murat; Hincal, A Atilla

2008-09-01

The objective of this study was to determine cyclosporine A (Cy A) levels in ocular tissues and fluids after topical administration of poly-epsilon-caprolactone (PCL)/benzalkonium chloride (BKC) nanospheres and hyaluronic acid (HA) coated PCL/BKC nanospheres onto healthy rabbit corneas. Nanospheres were prepared by nanoprecipitation and purified by gradient-rate centrifugation. Cy A (0.1%) in either castor oil solution (group 1), PCL/BKC nanosphere formulation (group 2) or HA coated PCL/BKC nanosphere formulation (group 3) was instilled onto rabbit corneas. Tear samples were adsorbed onto Schirmer tear strips. Cy A concentrations of fluid (blood, aqueous humor, tear) and specimen extracts (cornea, conjunctiva, iris/ciliary body) were determined by high performance liquid chromatography-mass spectrometry (LC-MS). The mean corneal Cy A concentration obtained at 0.5, 1, 2, 4, 8 and 24h following instillation of the formulations ranged between 0.12 and 1.2 ng/mg tissue for group 1, 5.9-15.5 ng/mg tissue for group 2 and 11.4-23.0 ng/mg for group 3 (one-way analysis of variance (ANOVA) and pairwise tests (SNK (Student-Newman-Keuls) and Tukey); p<0.05). Conjunctival Cy A levels of group 2 and 3 were not significantly different at any of the time points tested. However, there was a significant difference between Cy A concentration of castor oil formulation and that of PCL/BKC nanosphere formulation at 1 and 8h (p<0.05). The mean iris/ciliary body concentrations obtained with the three formulations were not significantly different at any time point with the exception of group 2 levels being higher than those of groups 1 and 3 at 1h (p<0.05). The lowest ocular tear Cy A concentrations (16-114 ng/ml) were found following the instillation of HA coated PCL/BKC nanoparticles (group 3) during the time period tested. Cy A loaded PCL/BKC and HA coated PCL/BKC nanospheres are able to achieve high levels of Cy A in the cornea that is 10-15-fold higher than that is achieved with Cy A solution in castor oil. Nanosphere formulation and HA may play an important role in delivering high levels of cyclosporine A into the cornea.

Compensation of corneal oblique astigmatism by internal optics: a theoretical analysis.

PubMed

Liu, Tao; Thibos, Larry N

2017-05-01

Oblique astigmatism is a prominent optical aberration of peripheral vision caused by oblique incidence of rays striking the refracting surfaces of the cornea and crystalline lens. We inquired whether oblique astigmatism from these two sources should be expected, theoretically, to have the same or opposite signs across the visual field at various states of accommodation. Oblique astigmatism was computed across the central visual field for a rotationally-symmetric schematic-eye using optical design software. Accommodative state was varied by altering the apical radius of curvature and separation of the biconvex lens's two aspheric surfaces in a manner consistent with published biometry. Oblique astigmatism was evaluated separately for the whole eye, the cornea, and the isolated lens over a wide range of surface curvatures and asphericity values associated with the accommodating lens. We also computed internal oblique astigmatism by subtracting corneal oblique astigmatism from whole-eye oblique astigmatism. A visual field map of oblique astigmatism for the cornea in the Navarro model follows the classic, textbook description of radially-oriented axes everywhere in the field. Despite large changes in surface properties during accommodation, intrinsic astigmatism of the isolated human lens for collimated light is also radially oriented and nearly independent of accommodation both in theory and in real eyes. However, the magnitude of ocular oblique astigmatism is smaller than that of the cornea alone, indicating partial compensation by the internal optics. This implies internal oblique astigmatism (which includes wavefront propagation from the posterior surface of the cornea to the anterior surface of the lens and intrinsic lens astigmatism) must have tangentially-oriented axes. This non-classical pattern of tangential axes for internal astigmatism was traced to the influence of corneal power on the angles of incidence of rays striking the internal lens. Partial compensation of corneal astigmatism by internal optics is due mainly to the highly converging nature of wavefronts incident upon the lens resulting from corneal refraction. The degree of compensation is quadratically dependent on eccentricity but is expected to diminish as the eye accommodates. Neutralising the cornea by index-matching defeats internal compensation, revealing classical, radially-oriented oblique astigmatism in the isolated lens. © 2017 The Authors Ophthalmic & Physiological Optics © 2017 The College of Optometrists.

Biomedical Research, Development, and Engineering at the Johns Hopkins University Applied Physics Laboratory Annual Report, October 1, 1975-September 30, 1976

DTIC Science & Technology

1976-09-01

Holographic Stress Testing of the surgicai. cornea, B. F. Hochheimer (APL) and J. .. Calkins (JHMI) §37 Constant Pressure Tonography, L. J. Viernatein...September 1976 HOLOGRAPHIC STRESS TESTING OF SUMMARY AND DISCUSSION THE SURGICAL CORNEA We have successfully demonstrated that holographic interferometry is...recording me- stress testing of corneal wounds in several thodology suitable for performing do sle-pulsed postoperative patients. The only stress used

Analysis of High Energy Laser Weapon Employment from a Navy Ship

DTIC Science & Technology

2012-09-01

meters Capacity ~ 400 kg (T.B. Racing and Marine, 2011) Speed 80 knots (148 km/h = 42 m/s) Material Fiberglass / Aluminium Explosive Improvised...CIWS) and requires the ship’s electrical power of about 400 kW. This would be deemed adequate for employment on an LCS ship. A boat target with an...Inflammation of the cornea, similar to sunburn 315 - 400 nm (Ultraviolet UV-A) CORNEA and LENS Photochemical cataract; Clouding of the lens 400

Adaptive cornea modeling from keratometric data.

PubMed

Martínez-Finkelshtein, Andrei; López, Darío Ramos; Castro, Gracia M; Alió, Jorge L

2011-07-01

To introduce an iterative, multiscale procedure that allows for better reconstruction of the shape of the anterior surface of the cornea from altimetric data collected by a corneal topographer. The report describes, first, an adaptive, multiscale mathematical algorithm for the parsimonious fit of the corneal surface data that adapts the number of functions used in the reconstruction to the conditions of each cornea. The method also implements a dynamic selection of the parameters and the management of noise. Then, several numerical experiments are performed, comparing it with the results obtained by the standard Zernike-based procedure. The numerical experiments showed that the algorithm exhibits steady exponential error decay, independent of the level of aberration of the cornea. The complexity of each anisotropic Gaussian-basis function in the functional representation is the same, but the parameters vary to fit the current scale. This scale is determined only by the residual errors and not by the number of the iteration. Finally, the position and clustering of the centers, as well as the size of the shape parameters, provides additional spatial information about the regions of higher irregularity. The methodology can be used for the real-time reconstruction of both altimetric data and corneal power maps from the data collected by keratoscopes, such as the Placido ring-based topographers, that will be decisive in early detection of corneal diseases such as keratoconus.

Translational Immuno- and Neuro-imaging Demonstrate Corneal Neuro-immune Crosstalk

PubMed Central

Hamrah, Pedram; Seyed-Razavi, Yashar; Yamaguchi, Takefumi

2017-01-01

Corneal immuno- and neuro-imaging approaches facilitate in vivo analyses of the cornea, including high-resolution imaging of corneal immune cells and nerves. This approach facilitates the analyses of underlying immune and nerve alterations not detected by clinical slit-lamp examination alone. In this review, we describe recent work performed in our translational ocular immunology center with a focus on ‘bench-to-bedside’ and ‘bedside-to-bench’ research. The ability to visualize dendritiform immune cells (DCs) in patients with laser in vivo confocal microscopy (IVCM), recently discovered in the central murine cornea, has allowed us to demonstrated their utility as a potential surrogate biomarker for inflammatory ocular surface diseases. This biomarker for inflammation allows the measurement of therapeutic efficacy of anti-inflammatory drugs and its utility as an endpoint in clinical trials with high inter-observer agreement. IVCM image analyses from our studies demonstrated a significant increase in DC density and size in ocular disease, a positive correlation between DC density and clinical signs and symptoms of disease and pro-inflammatory tear cytokines, and a strong negative correlation between DC density and subbasal nerve density. In conjunction with pre-clinical research investigating the inflammatory state in a partial or fully denervated cornea, our results indicated that corneal nerves are directly involved in the regulation of homeostasis and immune privilege in the cornea. PMID:27631352

Experimental results of a new system using microwaves for vision correction

NASA Astrophysics Data System (ADS)

Ryan, Thomas P.; Pertaub, Radha; Meyers, Steven R.; Dresher, Russell P.; Scharf, Ronald

2009-02-01

Technology is in development to correct vision without the use of lasers or cutting of the eye. Many current technologies used to reshape the cornea are invasive, in that either RF needles are placed into the cornea or a flap is cut and then a laser used to ablate the cornea in the optical zone. Keraflex, a therapeutic microwave treatment, is a noninvasive, non-incisional refractive surgery procedure capable of treating myopia (nearsightedness). The goal is to create a predictable refractive change in the optical zone, while preserving the epithelium and deeper structures of the eye. A further goal is to avoid incisions and damage to the epithelium which both require a post-treatment healing period. Experimental work with fresh porcine eyes examined the following variables: duration of the RF pulse, RF power level, coolant amount and timing, electrode spacing, applanation force against the eye, initial eye temperature, and age of eye. We measured curvature changes of the eye with topography, Scheimpflug, Wavefront aberrometry or other means to characterize diopter change as an important endpoint. Other assessment includes evaluation of a fine white ring seen in the cornea following treatment. Dose studies have been done to correlate the treated region with energy delivered. The timing and dosing of energy and cooling were investigated to achieve the target diopter change in vision.

Biocompatibility and light transmission of liposomal lenses.

PubMed

Danion, Anne; Doillon, Charles J; Giasson, Claude J; Djouahra, Saliha; Sauvageau, Patrick; Paradis, Renée; Vermette, Patrick

2007-10-01

To validate the biocompatibility and transmittance properties of contact lenses bearing intact liposomes. These liposomal lenses loaded with therapeutics can be used as ophthalmic drug delivery systems. The biocompatibility of soft contact lenses, coated with liposomes was evaluated through in vitro direct and indirect cytocompatibility assays on human corneal epithelial cells, on reconstructed human corneas and on ex vivo rabbit corneas. The direct and indirect transmission spectra of liposome-covered lenses were also evaluated to test if they transmit all wavelengths of the ultraviolet-visible spectrum, to thereby fulfill their optical function, without gross alteration of the colors perception and with a minimum of light dispersion. Contact lenses bearing layers of stable liposomes did not induce any significant changes in cell viability and in cell growth, compared with lenses bearing no liposome. Elution assays revealed that no cytotoxic compound leaks from the lenses whether bearing liposomes or not. Histological analyses of reconstructed human corneas and ex vivo rabbit corneas directly exposed to liposomal lenses revealed neither alteration to the cell nor to the tissue structures. Contact lenses bearing layers of liposomes did not significantly affect light transmission compared with control lenses without liposome at the wavelength of maximal photopic sensitivity, i.e., 550 nm. In addition, the contact lenses afford more eye protection in the ultraviolet spectrum, compared with the control lenses. Liposomal contact lenses are biocompatible and their transmittance properties are not affected in the visible light range.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Ying, E-mail: yingliu@doheny.org; Sun Yet-sen University, Zhongshan Ophthalmic Center, State Key Ophthalmic Laboratory, Guangzhou 510060; Kawai, Kirio

Research highlights: {yields} Inactivation of Smad4 caused disruption in the development of the anterior segment. {yields} Inactivation of Smad4 failed to disrupt early lens development. {yields} Smad4 controlled lens cell cycle and cell death processes. {yields} Smad4 may regulate actin stress fiber assembly and eyelid epithelial movement. -- Abstract: Purpose: Signaling by members of the TGF{beta} superfamily of molecules is essential for embryonic development and homeostasis. Smad4, a key intracellular mediator in TGF{beta} signaling, forms transcriptional activator complexes with Activin-, BMP-, and TGF{beta}-restricted Smad proteins. However, the functional role of Smad4 in controlling different visual system compartments has not beenmore » fully investigated. Methods: Using the Pax6 promoter-driven Cre transgenic, smad4 was conditionally inactivated in the lens, cornea and ectoderm of the eyelids. Standard histological and molecular analytical approaches were employed to reveal morphological and cellular changes. Results: Inactivation of Smad4 in the lens led to microphthalmia and cataract formation in addition to the persistent adhesion of the retina to the lens and the iris to the cornea. Inactivation of Smad4 from the ectoderm of the eyelid and cornea caused disruption to eyelid fusion and proper development of the corneal epithelium and corneal stroma. Conclusions: Smad4 is required for the development and maintenance of the lens in addition to the proper development of the cornea, eyelids, and retina.« less

Quantifying the effects of hydration on corneal stiffness with optical coherence elastography

NASA Astrophysics Data System (ADS)

Singh, Manmohan; Li, Jiasong; Han, Zhaolong; Vantipalli, Srilatha; Aglyamov, Salavat R.; Twa, Michael D.; Larin, Kirill V.

2018-02-01

Several methods have been proposed to assess changes in corneal biomechanical properties due to various factors, such as degenerative diseases, intraocular pressure, and therapeutic interventions (e.g. corneal collagen crosslinking). However, the effect of the corneal tissue hydration state on corneal stiffness is not well understood. In this work, we induce low amplitude (< 10 μm) elastic waves with a focused micro air-pulse in fresh in situ rabbit corneas (n = 10) in the whole eye-globe configuration at an artificially controlled intraocular pressure. The waves were then detected with a phase-stabilized swept source optical coherence elastography system. Baseline measurements were taken every 20 minutes for an hour while the corneas were hydrated with 1X PBS. After the measurement at 60 minutes, a 20% dextran solution was topically instilled to dehydrate the corneas. The measurements were repeated every 20 minutes again for an hour. The results showed that the elastic wave velocity decreased as the corneal thickness decreased. Finite element modeling (FEM) was performed using the corneal geometry and elastic wave propagation speed to assess the stiffness of the samples. The results show that the stiffness increased from 430 kPa during hydration with PBS to 500 kPa after dehydration with dextran, demonstrating that corneal hydration state, apart from geometry and intraocular pressure, can change the stiffness of the cornea.

The effect of corneal anterior surface eccentricity on astigmatism after cataract surgery.

PubMed

Park, Choul Yong; Chuck, Roy S; Channa, Prabjot; Lim, Chi-Yeon; Ahn, Byung-Jin

2011-01-01

To evaluate the effect of cornea eccentricity on induced astigmatism after cataract surgery. The study included 125 eyes of 87 patients. Preoperative corneal astigmatism, pachymetry, and eccentricity were measured. During cataract surgery, the location of the main incision (2.8-mm clear corneal) was selected to be either superior, superior-nasal, superior-temporal, nasal, or temporal to decrease the preexisting corneal astigmatism. Aspheric intraocular lenses were implanted. Keratometry and manifest refraction were recorded 6 months after surgery. Astigmatism was calculated using vector subtraction software. Three parameters significantly affected postoperative astigmatism: preoperative amount of corneal astigmatism, eccentricity of anterior cornea, and location of the main incision. The mean surgically induced astigmatism (SIA) was calculated to be: superior = 0.82 diopters (D), superior-nasal = 0.50 D, superior-temporal = 0.63 D, temporal = 0.45 D, and nasal = 0.55 D. Superior incision induced the greatest SIA and temporal incision induced the smallest SIA. The eccentricity of anterior cornea showed significantly positive correlation with the amount of SIA (P < .001). The preoperative corneal cylinder power showed significantly positive correlation with the amount of SIA (P < .001). Postoperative astigmatism was affected by various factors in cataract surgery. The greatest postoperative astigmatism is expected in corneas with high anterior eccentricity, high preoperative corneal astigmatism, and superior location of the main incision. Copyright 2011, SLACK Incorporated.

Corneal Thickness Profile and Associations in Chinese Children Aged 7 to 15 Years Old.

PubMed

Ma, Yingyan; Zhu, Xiaofeng; He, Xiangui; Lu, Lina; Zhu, Jianfeng; Zou, Haidong

2016-01-01

Corneal thickness (CT) maps of the central (2-mm diameter), para-central (2 to 5-mm diameter), peripheral (5 to 6-mm diameter), and minimum (5-mm diameter) cornea were measured in normal Chinese school children aged 7 to 15 years old using Fourier-domain optical coherence tomography. Multiple regression analyses were performed to explore the effect of associated factors [age, gender, refraction, axial length and corneal curvature radius (CCR)] on CT and the relationship between central corneal thickness (CCT) and intraocular pressure (IOP). A total of 1228 eyes from 614 children were analyzed. The average CCT was 532.96 ± 28.33 μm for right eyes and 532.70 ±28.45 μm for left eyes. With a 10 μm increase in CCT, the IOP was elevated by 0.37 mm Hg, as measured by noncontact tonometry. The CT increased gradually from the center to the periphery. The superior and superior nasal regions had the thickest CTs, while the thinnest points were primarily located in the inferior temporal cornea. The CCT was associated with CCR (p = 0.008) but not with gender (p = 0.075), age (p = 0.286), axial length (p = 0.405), or refraction (p = 0.985). In the para-central region and the peripheral cornea, increased CT was associated with younger age, male gender, and a flatter cornea.

Effect of marking pens on femtosecond laser-assisted flap creation.

PubMed

Ide, Takeshi; Kymionis, George D; Abbey, Ashkan M; Yoo, Sonia H; Culbertson, William W; O'Brien, Terrence P

2009-06-01

To compare the ease of the flap lift after central corneal marking with 2 types of marking pens after femtosecond laser-assisted flap creation in laser in situ keratomileusis. Bascom Palmer Eye Institute, University of Miami, Miami, Florida, USA. Porcine eyes were prepared for flap creation with a femtosecond laser (IntraLase). The eyes were assigned to 1 of 4 groups. After the femtosecond laser treatment, the difficulty of flap lifting the 4 groups was compared. Twelve porcine eyes, 3 in each group, were evaluated. In the 2 groups in which an oil-based pen was used, the corneal flap could not be lifted. In the 2 groups in which a water-based pen was used, the corneal flap was easily lifted. Oil-based ink may reduce the ability of the femtosecond laser to penetrate the cornea. The resultant corneal flap may require aggressive manipulation to be lifted. When used to mark the center of the cornea before flap creation, water-based ink provided greater ease of corneal flap lifting than oil-based ink. Because the marking is located over the center of the pupil, any alteration of the cornea in this area from aggressive flap lifting may result in substantial visual loss. Therefore, the use of an oil-based ink to mark the central cornea must be avoided to prevent traumatic irregularities of the flap stroma.

Determination of corneal elasticity coefficient using the ORA database.

PubMed

Avetisov, Sergei E; Novikov, Ivan A; Bubnova, Irina A; Antonov, Alexei A; Siplivyi, Vladimir I

2010-07-01

To propose a new approach for the study of corneal biomechanics using the Reichert Ocular Response Analyzer (ORA) database, which is based on changes in velocity retardation in the central cornea at the peak of flattening. The ORA applanation curve was analyzed using a mathematical technique, which allowed calculation of the elasticity coefficient (Ke), which is primarily characteristic of the elastic properties of the cornea. Elasticity coefficient values were obtained in patients with presumably different biomechanical properties of the cornea: "normal" cornea (71 eyes, normal group), keratoconus (34 eyes, keratoconus group), LASIK (36 eyes, LASIK group), and glaucoma with elevated and compensated intraocular pressure (lOP) (38 eyes, glaucoma group). The mean Ke value in the normal group was 11.05 +/- 1.6, and the corneal thickness correlation coefficient r2 was 0.48. In the keratoconus group, the mean Ke value was 4.91 +/- 1.87 and the corneal thickness correlation coefficient r2 was 0.47. In the LASIK group, Ke and r2 were 5.99 +/- 1.18 and 0.39, respectively. In the glaucoma group, the same eyes that experienced a two-fold reduction in lOP developed a statistically significant reduction in the Ke (1.06 times lower), whereas their corneal hysteresis value increased 1.25 times. The elasticity coefficient calculated using the ORA applanation curve can be used in the evaluation of corneal biomechanical properties.

Femtosecond laser cutting of human corneas for the subbasal nerve plexus evaluation.

PubMed

Kowtharapu, B S; Marfurt, C; Hovakimyan, M; Will, F; Richter, H; Wree, A; Stachs, O; Guthoff, R F

2017-01-01

Assessment of various morphological parameters of the corneal subbasal nerve plexus is a valuable method of documenting the structural and presumably functional integrity of the corneal innervation in health and disease. The aim of this work is to establish a rapid, reliable and reproducible method for visualization of the human corneal SBP using femtosecond laser cut corneal tissue sections. Trephined healthy corneal buttons were fixed and processed using TissueSurgeon-a femtosecond laser based microtome, to obtain thick tissue sections of the corneal epithelium and anterior stroma cut parallel to the ocular surface within approximately 15 min. A near infrared femtosecond laser was focused on to the cornea approximately 70-90 μm from the anterior surface to induce material separation using TissueSurgeon. The obtained corneal sections were stained following standard immunohistochemical procedures with anti-neuronal β-III tubulin antibody for visualization of the corneal nerves. Sections that contained the epithelium and approximately 20-30 μm of anterior stroma yielded excellent visualisation of the SBP with minimal optical interference from underlying stromal nerves. In conclusion, the results of this study have demonstrated that femtosecond laser cutting of the human cornea offers greater speed, ease and reliability than standard tissue preparation methods for obtaining high quality thick sections of the anterior cornea cut parallel to the ocular surface. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea.

PubMed

Rossi, Francesca; Pini, Roberto; Menabuoni, Luca

2007-01-01

Corneal laser welding is a technique used clinically to induce the immediate sealing of corneal wounds. We present an experimental and model analysis of the temperature dynamics during diode laser-induced corneal welding, which is aimed at characterizing the mechanism of tissue fusion. Ex vivo tests were performed on porcine eyes in the typical irradiation conditions used for laser-induced suturing in cornea transplant. Three laser power densities (12.5 W/cm(2), 16.7 W/cm(2), 20.8 W/cm(2)) were tested. The superficial temperature of the cornea was measured by means of an infrared thermocamera. Experimental data were compared with the results of a three-dimensional (3D) model of a laser-welding process in the cornea, solved by the use of the Finite Element Method (FEM). The model solution and experimental results showed good agreement. The model was thus used to estimate the temperature enhancement inside the corneal wound and to calculate the thermal damage inside the tissue. The results indicated a selective, spatially confined heating effect that occurred at operative temperatures (59 to 66 degrees C) close to intermediate denaturation points of the stromal collagen, before its complete disorganization. No significant heat damage to the region of the laser-treated wound was evidenced in the operative irradiation conditions of corneal welding.

Safety of cornea and iris in ocular surgery with 355-nm lasers.

PubMed

Wang, Jenny; Chung, Jae Lim; Schuele, Georg; Vankov, Alexander; Dalal, Roopa; Wiltberger, Michael; Palanker, Daniel

2015-09-01

A recent study showed that 355-nm nanosecond lasers cut cornea with similar precision to infrared femtosecond lasers. However, use of ultraviolet wavelength requires precise assessment of ocular safety to determine the range of possible ophthalmic applications. In this study, the 355-nm nanosecond laser was evaluated for corneal and iris damage in rabbit, porcine, and human donor eyes as determined by minimum visible lesion (MVL) observation, live/dead staining of the endothelium, and apoptosis assay. Single-pulse damage to the iris was evaluated on porcine eyes using live/dead staining. In live rabbits, the cumulative median effective dose (ED50) for corneal damage was 231 J/cm2, as seen by lesion observation. Appearance of endothelial damage in live/dead staining or apoptosis occurred at higher radiant exposure of 287 J/cm2. On enucleated rabbit and porcine corneas, ED50 was 87 and 52 J/cm2, respectively, by MVL, and 241 and 160 J/cm2 for endothelial damage. In human eyes, ED50 for MVL was 110 J/cm2 and endothelial damage at 453 J/cm2. Single-pulse iris damage occurred at ED 50 of 208 mJ/cm2. These values determine the energy permitted for surgical patterns and can guide development of ophthalmic laser systems. Lower damage threshold in corneas of enucleated eyes versus live rabbits is noted for future safety evaluation.

The effects of sperminated pullulans on cornea permeability to puerarin and the toxicity.

PubMed

Yu, Nannan; Dong, Guo; Ge, Hongyan; Jin, Di; Cui, Hao; Liu, Ping

2012-10-01

To investigate the varied effects of sperminated pullulans (SP) with different amino residues on cornea permeability and its local toxicity. Three groups of rabbits were used: control, low-amino residue content SP (SP-L), and high-amino residue content SP (SP-H). The in vitro and in vivo spreading assays were combined with high performance liquid chromatography (HPLC) to measure the concentration of puerarin in the external medium or aqueous humor when 0% SP, 0.2% SP-L, and 0.2% SP-H were included. The toxicity of SP was determined by corneal hydration values, Draize score, aqueous humor protein concentration, corneal endothelial evaluation, as well as light microscopy and electron microscopy. The application of 0.2% SP-L and 0.2% SP-H to the cornea in vitro increased puerarin apparent permeability coefficient by 1.96-fold (P<0.05) and 2.95-fold (P<0.01), respectively. SP-H showed stronger effect than SP-L (P<0.05). For the in vivo assay, those were 1.81-fold (P<0.05) and 3.71-fold (P<0.01), respectively. With the SP application, the corneal hydration values were <83% and Draize scores were <4, with no apparent changes in histological observations. SP is one potential adjuvant promoting puerarin permeability to the cornea, and the high-content amino residue SP showed stronger effect, without ocular toxicity.

Dynamic behavior of photoablation products of corneal tissue in the mid-IR: a study with FELIX

NASA Astrophysics Data System (ADS)

Auerhammer, J. M.; Walker, R.; van der Meer, A. F. G.; Jean, B.

The properties of pulsed IR-laser ablation of biological soft tissue (porcine cornea) were studied in vitro systematically and quantitatively with a free-electron laser in the wavelength range 6<=λ<=20 μm at fluences ranging from 3.1 to 9.4 J/cm2. Dynamic parameters such as the extension of the ablation cloud, the initial velocity and momentum of the ablated particles as well as the ablation threshold, the ablated mass, and the particle size were investigated. The ablation plume was made visible with a stroboscopic technique. For a fluence of 3.1 J/cm2 the average initial velocity of the ejected particles was deduced from the extension of the plume to range from 120-400 m/s. Measurements of the recoil momentum using a sensitive pendulum led to values between 0.5 and 2.0 mmg/s. All measured properties were related to the spectroscopically determined absorption coefficient of cornea αcornea. Where absorption due to proteins is high (at λ=6.2 and 6.5 μm), ablated mass, velocity and recoil momentum behave according to αcornea. For the first time, variations of the ablation plume from pulse to pulse were observed. Those, as well as the particle size, not only depend on the absorption coefficient, but also on the predominant absorber.

Involvement of NADPH oxidases in alkali burn-induced corneal injury.

PubMed

Gu, Xue-Jun; Liu, Xian; Chen, Ying-Ying; Zhao, Yao; Xu, Man; Han, Xiao-Jian; Liu, Qiu-Ping; Yi, Jing-Lin; Li, Jing-Ming

2016-07-01

Chemical burns are a major cause of corneal injury. Oxidative stress, inflammatory responses and neovascularization after the chemical burn aggravate corneal damage, and lead to loss of vision. Although NADPH oxidases (Noxs) play a crucial role in the production of reactive oxygen species (ROS), the role of Noxs in chemical burn-induced corneal injury remains to be elucidated. In the present study, the transcription and expression of Noxs in corneas were examined by RT-qPCR, western blot analysis and immunofluorescence staining. It was found that alkali burns markedly upregulated the transcription and expression of Nox2 and Nox4 in human or mouse corneas. The inhibition of Noxs by diphenyleneiodonium (DPI) or apocynin (Apo) effectively attenuated alkali burn-induced ROS production and decreased 3-nitrotyrosine (3-NT) protein levels in the corneas. In addition, Noxs/CD11b double‑immunofluorescence staining indicated that Nox2 and Nox4 were partially co-localized with CD11b. DPI or Apo prevented the infiltration of CD11b-positive inflammatory cells, and inhibited the transcription of inflammatory cytokines following alkali burn-induced corneal injury. In our mouse model of alkali burn-induced corneal injury, corneal neovascularization (CNV) occurred on day 3, and it affected 50% of the whole area of the cornea on day 7, and on day 14, CNV coverage of the cornea reached maximum levels. DPI or Apo effectively attenuated alkali burn‑induced CNV and decreased the mRNA levels of angiogenic factors, including vascular endothelial growth factor (VEGF), VEGF receptors and matrix metalloproteinases (MMPs). Taken together, our data indicate that Noxs play a role in alkali burn-induced corneal injury by regulating oxidative stress, inflammatory responses and CNV, and we thus suggest that Noxs are a potential therapeutic target in the future treatment of chemical-induced corneal injury.

Anti-neovascular effect of chondrocyte-derived extracellular matrix on corneal alkaline burns in rabbits.

PubMed

Lee, Hye Sook; Lee, Ji Hyun; Kim, Chae Eun; Yang, Jae Wook

2014-06-01

We investigated the effect of a chondrocyte-derived extracellular matrix (CDECM) on experimental corneal alkaline burns in rabbits. Corneal neovascularization (NV) was induced by applying an 8-mm filter paper soaked in 1 N NaOH to the right central corneas of rabbits for 1 minute. Ten days later, the rabbits were randomly divided into three groups: the alkaline burn group, the CDECM transplantation group, and the human amniotic membrane (HAM) transplantation group. The left eyes were used as controls. CDECM and HAM were transplanted onto the corneal surface to completely cover the resected area and were subsequently sutured. On the 10th day after transplantation, the structural changes of the cornea were analyzed histologically. We examined the effects of CDECM on clinical NV features and on the expression of corneal NV markers. The alkaline burn produced significant NV and increased the corneal thickness. On day 10 after transplantation, the thickness, NV and opacity of the cornea were markedly decreased in the CDECM group (p < 0.001). However, the HAM transplantation group did not exhibit improvements in these clinical parameters, and there were no significant differences relative to the burn group. In addition, the use of CDECM improved the healing of the cornea following the alkaline burn by disrupting the corneal epithelial proliferation and reducing the fibrotic changes of the stroma. The hallmarks of NV were significantly induced in the subepithelium by the alkaline burn, and these levels were also suppressed by CDECM. The CDECM suppressed corneal NV by inhibiting nuclear factor-kappa B (NF-κB) activation by blocking the PKC and Akt signaling pathways. CDECM transplantation was markedly effective in healing alkali-burned corneas by modulating the translocation of NF-κB to the nucleus, thereby representing a promising material for the noninvasive treatment of ocular surface disease.

Glutathione-related enzymes and the eye.

PubMed

Ganea, Elena; Harding, John J

2006-01-01

Glutathione and the related enzymes belong to the defence system protecting the eye against chemical and oxidative stress. This review focuses on GSH and two key enzymes, glutathione reductase and glucose-6-phosphate dehydrogenase in lens, cornea, and retina. Lens contains a high concentration of reduced glutathione, which maintains the thiol groups in the reduced form. These contribute to lens complete transparency as well as to the transparent and refractive properties of the mammalian cornea, which are essential for proper image formation on the retina. In cornea, gluthatione also plays an important role in maintaining normal hydration level, and in protecting cellular membrane integrity. In retina, glutathione is distributed in the different types of retinal cells. Intracellular enzyme, glutathione reductase, involved in reducing the oxidized glutathione has been found at highest activity in human and primate lenses, as compared to other species. Besides the enzymes directly involved in maintaining the normal redox status of the cell, glucose-6-phosphate dehydrogenase which catalyzes the first reaction of the pentose phosphate pathway, plays a key role in protection of the eye against reactive oxygen species. Cornea has a high activity of the pentose phosphate pathway and glucose-6-phosphate dehydrogenase activity. Glycation, the non-enzymic reaction between a free amino group in proteins and a reducing sugar, slowly inactivates gluthathione-related and other enzymes. In addition, glutathione can be also glycated. The presence of glutathione, and of the related enzymes has been also reported in other parts of the eye, such as ciliary body and trabecular meshwork, suggesting that the same enzyme systems are present in all tissues of the eye to generate NADPH and to maintain gluthatione in the reduced form. Changes of glutathione and related enzymes activity in lens, cornea, retina and other eye tissues, occur with ageing, cataract, diabetes, irradiation and administration of some drugs.

Colony Stimulating Factor-1 Receptor Expressing Cells Infiltrating the Cornea Control Corneal Nerve Degeneration in Response to HSV-1 Infection

PubMed Central

Chucair-Elliott, Ana J.; Gurung, Hem R.; Carr, Meghan M.; Carr, Daniel J. J.

2017-01-01

Purpose Herpes simplex virus type-1 (HSV-1) is a leading cause of neurotrophic keratitis, characterized by decreased or absent corneal sensation due to damage to the sensory corneal innervation. We previously reported the elicited immune response to infection contributes to the mechanism of corneal nerve regression/damage during acute HSV-1 infection. Our aim is to further establish the involvement of infiltrated macrophages in the mechanism of nerve loss upon infection. Methods Macrophage Fas-Induced Apoptosis (MAFIA) transgenic C57BL/6 mice were systemically treated with AP20187 dimerizer or vehicle (VEH), and their corneas, lymph nodes, and blood were assessed for CD45+CD11b+GFP+ cell depletion by flow cytometry (FC). Mice were ocularly infected with HSV-1 or left uninfected. At 2, 4, and/or 6 days post infection (PI), corneas were assessed for sensitivity and harvested for FC, nerve structure by immunohistochemistry, viral content by plaque assay, soluble factor content by suspension array, and activation of signaling pathways by Western blot analysis. C57BL6 mice were used to compare to the MAFIA mouse model. Results MAFIA mice treated with AP20187 had efficient depletion of CD45+CD11b+GFP+ cells in the tissues analyzed. The reduction of CD45+CD11b+GFP+ cells recruited to the infected corneas of AP20187-treated mice correlated with preservation of corneal nerve structure and function, decreased protein concentration of inflammatory cytokines, and decreased STAT3 activation despite no changes in viral content in the cornea compared to VEH-treated animals. Conclusions Our results suggest infiltrated macrophages are early effectors in the nerve regression following HSV-1 infection. We propose the neurodegeneration mechanism involves macrophages, local up-regulation of IL-6, and activation of STAT3. PMID:28903153

The impact of intraocular pressure on elastic wave velocity estimates in the crystalline lens.

PubMed

Park, Suhyun; Yoon, Heechul; Larin, Kirill V; Emelianov, Stanislav Y; Aglyamov, Salavat R

2016-12-20

Intraocular pressure (IOP) is believed to influence the mechanical properties of ocular tissues including cornea and sclera. The elastic properties of the crystalline lens have been mainly investigated with regard to presbyopia, the age-related loss of accommodation power of the eye. However, the relationship between the elastic properties of the lens and IOP remains to be established. The objective of this study is to measure the elastic wave velocity, which represents the mechanical properties of tissue, in the crystalline lens ex vivo in response to changes in IOP. The elastic wave velocities in the cornea and lens from seven enucleated bovine globe samples were estimated using ultrasound shear wave elasticity imaging. To generate and then image the elastic wave propagation, an ultrasound imaging system was used to transmit a 600 µs pushing pulse at 4.5 MHz center frequency and to acquire ultrasound tracking frames at 6 kHz frame rate. The pushing beams were separately applied to the cornea and lens. IOP in the eyeballs was varied from 5 to 50 mmHg. The results indicate that while the elastic wave velocity in the cornea increased from 0.96  ±  0.30 m s -1 to 6.27  ±  0.75 m s -1 as IOP was elevated from 5 to 50 mmHg, there were insignificant changes in the elastic wave velocity in the crystalline lens with the minimum and the maximum speeds of 1.44  ±  0.27 m s -1 and 2.03  ±  0.46 m s -1 , respectively. This study shows that ultrasound shear wave elasticity imaging can be used to assess the biomechanical properties of the crystalline lens noninvasively. Also, it was observed that the dependency of the crystalline lens stiffness on the IOP was significantly lower in comparison with that of cornea.

Corneal biomechanical properties from two-dimensional corneal flap extensiometry: application to UV-riboflavin cross-linking.

PubMed

Kling, Sabine; Ginis, Harilaos; Marcos, Susana

2012-07-27

Corneal biomechanical properties are usually measured by strip extensiometry or inflation methods. We developed a two-dimensional (2D) flap extensiometry technique, combining the advantages of both methods, and applied it to measure the effect of UV-Riboflavin cross-linking (CXL). Corneal flaps (13 pig/8 rabbit) from the de-epithelialized anterior stroma (96 μm) were mounted on a custom chamber, consisting of a BK7 lens, a reflective retina, and two reservoirs (filled with Riboflavin and silicone oil). Stretching the corneal flap during five pressure increase/decrease cycles (0-30 mm Hg) changed the refractive power of the system, whose Zernike aberrations were monitored with a ray-tracing aberrometer. Porcine flaps were used to test the system. Rabbits were treated with CXL unilaterally in vivo following standard clinical procedures. Flaps were measured 1 month postoperatively. An analytical model allowed estimating Young's modulus from the change in surface (strain) and pressure (stress). Confocal microscopy examination was performed before, and at different times after CXL. Flap curvature changed with increased function of IOP in pig flaps (23.4 × 10⁻³ D/mm Hg). In rabbit flaps curvature changed significantly less in 1 month post CXL (P = 0.026) than in untreated corneas [17.0 vs. 6.36 millidiopter (mD)/mm Hg]. Young's modulus was 2.29 megapascals (MPa) in porcine corneas, 1.98 MPa in untreated rabbit corneas, and 4.83 MPa in 1 month post CXL rabbit corneas. At the same time, highly reflective structures were observed in the rabbit midstroma after treatment. 2D flap extensiometry allows estimating corneal elasticity in vitro. The measurements are spatially resolved in depth, minimize the effects of corneal hydration, and preserve the integrity of the cornea. The method proved the efficacy of CXL in increasing corneal rigidity after 1 month in rabbits.

Determining the Depth of Injury in Bioengineered Tissue Models of Cornea and Conjunctiva for the Prediction of All Three Ocular GHS Categories

PubMed Central

Zorn-Kruppa, Michaela; Houdek, Pia; Wladykowski, Ewa; Engelke, Maria; Bartok, Melinda; Mewes, Karsten R.; Moll, Ingrid; Brandner, Johanna M.

2014-01-01

The depth of injury (DOI) is a mechanistic correlate to the ocular irritation response. Attempts to quantitatively determine the DOI in alternative tests have been limited to ex vivo animal eyes by fluorescent staining for biomarkers of cell death and viability in histological cross sections. It was the purpose of this study to assess whether DOI could also be measured by means of cell viability detected by the MTT assay using 3-dimensional (3D) reconstructed models of cornea and conjunctiva. The formazan-free area of metabolically inactive cells in the tissue after topical substance application is used as the visible correlate of the DOI. Areas of metabolically active or inactive cells are quantitatively analyzed on cryosection images with ImageJ software analysis tools. By incorporating the total tissue thickness, the relative MTT-DOI (rMTT-DOI) was calculated. Using the rMTT-DOI and human reconstructed cornea equivalents, we developed a prediction model based on suitable viability cut-off values. We tested 25 chemicals that cover the whole range of eye irritation potential based on the globally harmonized system of classification and labelling of chemicals (GHS). Principally, the MTT-DOI test method allows distinguishing between the cytotoxic effects of the different chemicals in accordance with all 3 GHS categories for eye irritation. Although the prediction model is slightly over-predictive with respect to non-irritants, it promises to be highly valuable to discriminate between severe irritants (Cat. 1), and mild to moderate irritants (Cat. 2). We also tested 3D conjunctiva models with the aim to specifically address conjunctiva-damaging substances. Using the MTT-DOI method in this model delivers comparable results as the cornea model, but does not add additional information. However, the MTT-DOI method using reconstructed cornea models already provided good predictability that was superior to the already existing established in vitro/ex vivo methods. PMID:25494045

The effect of riboflavin/UVA collagen cross-linking therapy on the structure and hydrodynamic behaviour of the ungulate and rabbit corneal stroma.

PubMed

Hayes, Sally; Kamma-Lorger, Christina S; Boote, Craig; Young, Robert D; Quantock, Andrew J; Rost, Anika; Khatib, Yasmeen; Harris, Jonathan; Yagi, Naoto; Terrill, Nicholas; Meek, Keith M

2013-01-01

To examine the effect of riboflavin/UVA corneal crosslinking on stromal ultrastructure and hydrodynamic behaviour. One hundred and seventeen enucleated ungulate eyes (112 pig and 5 sheep) and 3 pairs of rabbit eyes, with corneal epithelium removed, were divided into four treatment groups: Group 1 (28 pig, 2 sheep and 3 rabbits) were untreated; Group 2 (24 pig) were exposed to UVA light (3.04 mW/cm(2)) for 30 minutes and Group 3 (29 pig) and Group 4 (31 pig, 3 sheep and 3 rabbits) had riboflavin eye drops applied to the corneal surface every 5 minutes for 35 minutes. Five minutes after the initial riboflavin instillation, the corneas in Group 4 experienced a 30 minute exposure to UVA light (3.04 mW/cm(2)). X-ray scattering was used to obtain measurements of collagen interfibrillar spacing, spatial order, fibril diameter, D-periodicity and intermolecular spacing throughout the whole tissue thickness and as a function of tissue depth in the treated and untreated corneas. The effect of each treatment on the hydrodynamic behaviour of the cornea (its ability to swell in saline solution) and its resistance to enzymatic digestion were assessed using in vitro laboratory techniques. Corneal thickness decreased significantly following riboflavin application (p<0.01) and also to a lesser extent after UVA exposure (p<0.05). With the exception of the spatial order factor, which was higher in Group 4 than Group 1 (p<0.01), all other measured collagen parameters were unaltered by cross-linking, even within the most anterior 300 microns of the cornea. The cross-linking treatment had no effect on the hydrodynamic behaviour of the cornea but did cause a significant increase in its resistance to enzymatic digestion. It seems likely that cross-links formed during riboflavin/UVA therapy occur predominantly at the collagen fibril surface and in the protein network surrounding the collagen.

The Effect of Riboflavin/UVA Collagen Cross-linking Therapy on the Structure and Hydrodynamic Behaviour of the Ungulate and Rabbit Corneal Stroma

PubMed Central

Hayes, Sally; Kamma-Lorger, Christina S.; Boote, Craig; Young, Robert D.; Quantock, Andrew J.; Rost, Anika; Khatib, Yasmeen; Harris, Jonathan; Yagi, Naoto; Terrill, Nicholas; Meek, Keith M.

2013-01-01

Purpose To examine the effect of riboflavin/UVA corneal crosslinking on stromal ultrastructure and hydrodynamic behaviour. Methods One hundred and seventeen enucleated ungulate eyes (112 pig and 5 sheep) and 3 pairs of rabbit eyes, with corneal epithelium removed, were divided into four treatment groups: Group 1 (28 pig, 2 sheep and 3 rabbits) were untreated; Group 2 (24 pig) were exposed to UVA light (3.04 mW/cm2) for 30 minutes and Group 3 (29 pig) and Group 4 (31 pig, 3 sheep and 3 rabbits) had riboflavin eye drops applied to the corneal surface every 5 minutes for 35 minutes. Five minutes after the initial riboflavin instillation, the corneas in Group 4 experienced a 30 minute exposure to UVA light (3.04 mW/cm2). X-ray scattering was used to obtain measurements of collagen interfibrillar spacing, spatial order, fibril diameter, D-periodicity and intermolecular spacing throughout the whole tissue thickness and as a function of tissue depth in the treated and untreated corneas. The effect of each treatment on the hydrodynamic behaviour of the cornea (its ability to swell in saline solution) and its resistance to enzymatic digestion were assessed using in vitro laboratory techniques. Results Corneal thickness decreased significantly following riboflavin application (p<0.01) and also to a lesser extent after UVA exposure (p<0.05). With the exception of the spatial order factor, which was higher in Group 4 than Group 1 (p<0.01), all other measured collagen parameters were unaltered by cross-linking, even within the most anterior 300 microns of the cornea. The cross-linking treatment had no effect on the hydrodynamic behaviour of the cornea but did cause a significant increase in its resistance to enzymatic digestion. Conclusions It seems likely that cross-links formed during riboflavin/UVA therapy occur predominantly at the collagen fibril surface and in the protein network surrounding the collagen. PMID:23349690

Regeneration of defective epithelial basement membrane and restoration of corneal transparency

PubMed Central

Marino, Gustavo K.; Santhiago, Marcony R.; Santhanam, Abirami; Torricelli, Andre A. M.; Wilson, Steven E.

2018-01-01

PURPOSE To study regeneration of the normal ultrastructure of the epithelial basement membrane (EBM) in rabbit corneas that had -9D photorefractive keratectomy (PRK) and developed late haze (fibrosis) with restoration of transparency over one to four months after surgery and in corneas that had incisional wounds. METHODS Twenty-four rabbits had one of their eyes included into one of the two procedure groups (-9D PRK or nearly full-thickness incisional wounds), while the opposite eye serving as unwounded controls. All corneas were evaluated with slit lamp photos, transmission electron microscopy and immunohistochemistry for the myofibroblast marker alpha-smooth muscle actin and collagen type III. RESULTS In the ‘-9D PRK group’, corneas at one month after surgery had dense corneal haze and no evidence of regenerated EBM ultrastructure. By two months after surgery, however, small areas of stromal clearing began to appear within the confluent opacity (lacunae), and these corresponded to small islands of normally-regenerated EBM detected within larger area of the excimer laser-ablated zone with no evidence of normal EBM. By four months after surgery, the EBM was fully-regenerated and the corneal transparency was completely restored to the ablated zone. In the ‘Incisional wound group’, the two dense, linear corneal opacities were observed at one month after surgery and progressively faded by two and three months after surgery. The EBM ultrastructure was fully regenerated at the site of the incisions, including around epithelial plugs that extended into the stroma, by one month after surgery in all eyes. CONCLUSIONS In the rabbit model, spontaneous resolution of corneal fibrosis (haze) after high correction PRK is triggered by regeneration of EBM with normal ultrastructure in the excimer laser- ablated zone. Conversely, incisional wounds heal in rabbit corneas without the development of myofibroblasts because the EBM regenerates normally by one month after surgery. PMID:28486725

[To Protect Corneal Transparency against Diseases].

PubMed

Usui, Tomohiko

2016-03-01

To protect corneal transparency, we tried to develop a new therapeutic strategy for corneal neovascularization, corneal scar, and TGFBI-related corneal dystrophy using nucleic acid drug. 1. The expression of angiopietin-like protein 2 (Angptl2) markedly increased in the neovascularized corneas compared to the normal cornea, and Angtpl2 was(a potent inducer of inflammatory corneal neovascularization. We have produced a single-stranded proline-modified short hairpin anti-Angptl2 ribonucleric acid interference (RNAi) molecule that is carried in a lipid nanoparticle for topical application. We have found this agent can penetrate all layers of the cornea. Angptl2 mRNA expression and corneal neovascularization were inhibited in a mouse alkari injury model by topical application of this agent. Thus, this modified RNAi agent is a new topical formulation for use against corneal neovascularization and scar. 2. Human umbilical vein endothelial cells (HUVECs) were cultured with human corneal keratocytes under serum-free conditions. We performed microarray gene-expression analysis in the coculture system and selected angiopoietin-like protein 7 (Angptl7). In vivo, intrastromal injections of an anti-Angptl7 RNAi agent into the avascular corneal stroma of mice resulted in the growth of blood vessels. Further, we examined the effects of Angptl7 on corneal nerves using culture rat trigeminal cells and this molecule had neurotrophic property on the cornea. Thus, Angpt17 is a unique molecule, which contain its bilateral character (anti-angiogenic and neurotrophic) in the cornea; an agonistic nucleic acid drug for Angptl7 may be a new therapeutic tool for protecting corneal transparency. 3. We examined local gene editing for TGFBI-related corneal dystrophy using CRISPR-Cas9 mediated homology directed repair (HDR). Cultured corneal keratocytes were obtained from a patient of R124H granular dystrophy. The R124H gene arrangement was corrected by a tranfection of guide RNA and HDR repair template single strand DNA in vitro. Thus, CRISPR-Cas9 medi-ated HDR could be a future radical treatment for TGFBI-related corneal dystrophy.

Fibroblast growth factor receptor 2 (FGFR2) is required for corneal epithelial cell proliferation and differentiation during embryonic development.

PubMed

Zhang, Jinglin; Upadhya, Dinesh; Lu, Lin; Reneker, Lixing W

2015-01-01

Fibroblast growth factors (FGFs) play important roles in many aspects of embryonic development. During eye development, the lens and corneal epithelium are derived from the same surface ectodermal tissue. FGF receptor (FGFR)-signaling is essential for lens cell differentiation and survival, but its role in corneal development has not been fully investigated. In this study, we examined the corneal defects in Fgfr2 conditional knockout mice in which Cre expression is activated at lens induction stage by Pax6 P0 promoter. The cornea in LeCre, Fgfr2(loxP/loxP) mice (referred as Fgfr2(CKO)) was analyzed to assess changes in cell proliferation, differentiation and survival. We found that Fgfr2(CKO) cornea was much thinner in epithelial and stromal layer when compared to WT cornea. At embryonic day 12.5-13.5 (E12.5-13.5) shortly after the lens vesicle detaches from the overlying surface ectoderm, cell proliferation (judged by labeling indices of Ki-67, BrdU and phospho-histone H3) was significantly reduced in corneal epithelium in Fgfr2(CKO) mice. At later stage, cell differentiation markers for corneal epithelium and underlying stromal mesenchyme, keratin-12 and keratocan respectively, were not expressed in Fgfr2(CKO) cornea. Furthermore, Pax6, a transcription factor essential for eye development, was not present in the Fgfr2(CKO) mutant corneal epithelial at E16.5 but was expressed normally at E12.5, suggesting that FGFR2-signaling is required for maintaining Pax6 expression in this tissue. Interestingly, the role of FGFR2 in corneal epithelial development is independent of ERK1/2-signaling. In contrast to the lens, FGFR2 is not required for cell survival in cornea. This study demonstrates for the first time that FGFR2 plays an essential role in controlling cell proliferation and differentiation, and maintaining Pax6 levels in corneal epithelium via ERK-independent pathways during embryonic development.

[The relationship between corneal lymphangiogenesis and inflammation index after corneal alkali injury].

PubMed

Ling, Shi-qi; Li, Wei-hua; Xu, Jian-gang; Kuang, Wen-hui; Li, Chao-yang

2010-11-01

To discuss the relationship between corneal lymphangiogenesis and inflammation index (IF) in alkali burned corneas. Experimental research. Rat corneal hemangiogenesis and lymphangiogenesis were examined by 5'-nase-alkaline phosphatase (5'-NA-ALP) double enzyme-histochemistry and whole mount immunofluorescence at 1 day, 3 days, and 1, 2, 3, 4, 5, 6, 7, 8 weeks after alkaline burns, and the blood vessel counting (BVC) and the lymphatic vessel counting (LVC) were recorded. The state of corneal inflammation was observed under the slit lamp and evaluated by inflammation index (IF) grading at the same time. Then, the association of LVC with IF was examined. In addition, eleven human alkali burned corneas were obtained from 11 patients undergoing corneal transplantation in Zhongshan Ophthalmic Center from January 2005 to June 2008. Corneal lymphangiogenesis was examined by lymphatic vessel endothelial receptor (LYVE-1) immunohistochemistry. The significance of the differences in IF, inflammatory cells counting, burn history, and age between two groups was analyzed by using paired student's t-test. New lymphatic vessels were present in rat alkali burned corneas. Corneal lymphangiogenesis developed 3 days after alkaline burns, reached the top 2 weeks after the injury, then decreased gradually, and disappeared at the end of the 5th week. Corneal lymphatics occurred behind corneal inflammation, but disappeared before corneal inflammation and hemangiogenesis. LVC was strongly and positively correlated with IF (r = 0.572, P < 0.01) after corneal alkaline burns. Among eleven human alkali burned corneas, corneal lymphatic vessels were present in 3 corneas. Compared with the other 8 cases without corneal lymphangiogenesis, the scores of IF was significantly higher (t = 3.28, P < 0.05), the inflammatory cells counting dramatically increased (t = 2.42, P < 0.05), but the age decreased significantly (t = 2.62, P < 0.05). However, the difference in burn history between two groups was not significant (t = 1.28, P > 0.05). Corneal lymphangiogenesis develops after alkaline-burns and correlates closely with inflammation index.

[The glaucoma pharmacological treatment and biomechanical properties of the cornea].

PubMed

Liehneová, I; Karlovská, S

2014-10-01

To evaluate and compare the impact of long-term use of intraocular pressure lowering medication on the biomechanical properties of the cornea. Group of 305 eyes of 154 patients newly diagnosed with primary open angle glaucoma (POAG, n = 68) or ocular hypertension (OH, n = 6) was enrolled in prospective cohort study. The control group was established of 80 untreated eyes of 40 patients with ocular hypertension and 80 eyes of 40 patients with no ocular pathology. Following parameters were evaluated: intraocular pressure (IOPg,IOPcc), hysteresis (CH), corneal resistance factor (CRF) and central corneal thickness (CCT). The parameters were evaluated at baseline (untreated) and in follow up periods of 3, 6, 9 and 12 months. The same schedule was used for eyes in the control group. Eyes with POAG or OH were sorted into two groups depending on the type of applied medication: prostaglandin analogues, carboanhydrase inhibitors alone or combined with betablockers. We did not prove any statistically significant difference in hysteresis in patients with newly diagnosed POAG (yet untreated) in comparison with normal eyes in control group (p = 0.238). We proved significantly higher values of CRF (p = 0.032) and CCT (p = 0.013) in the control group of untreated patients with ocular hypertension. This result confirms higher number of patients with stiffer and thicker corneas. Statistically significant difference of CH and CRF was proved (p < 0.0001) in eyes treated by prostaglandin analogues during follow up period. In these eyes we also demonstrated reduction of CCT (p < 0.001). We did not record any other statistically significant change in remaining followed parameters. Increase of CH and CRF can show change of biomechanical properties of the cornea after long-term use of prostaglandin analogues. The biomechanical properties of the cornea were not impacted by carboanhydrase inhibitors. Further studies are required to establish the effect of long-term use prostaglandin analogues on accuracy of IOP measurements.

The eicosanoid, 15-(S)-HETE, stimulates secretion of mucin-like glycoprotein by the corneal epithelium.

PubMed

Jackson , R S; Van Dyken, S J; McCartney, M D; Ubels, J L

2001-07-01

The eicosanoid, 15-(S)-hydroxyeicosa-5Z, 8Z-11Z, 13E-tetraenoic acid (15-(S)-HETE), is known to stimulate production of mucin glycoprotein by airway epithelium. This study investigated the effect of 15-(S)-HETE on the mucin glycoprotein secretion by the corneal epithelium. To determine the effect of dose, corneas of anesthetized New Zealand White rabbits were treated with 50, 500, or 5,000 nM 15-(S)-HETE in artificial tears for 120 minutes. To determine the time to onset of the response, corneas were treated with 500 or 1,000 nM 15-(S)-HETE in balanced salt solution for periods ranging from 5 to 120 minutes. Corneas were fixed for electron microscopy in fixative containing 0.5% cetylpyridinium chloride (CPC) to stabilize the layer of mucin-like glycoprotein on the corneal surface. The mucin layer thickness was measured by image analysis of electron micrographs. The layer of CPC-fixed mucin-like glycoprotein on the surface of control corneas was 0.46 +/- 0.04 microm thick. After treatment with 15-(S)-HETE, the thickness of the mucin layer increased to 0.64 +/- 0.1 microm at 50 or 5,000 nM HETE and as much as 1.02 +/- 0.2 microm in response to 500 nM HETE. Mucin thickness reached a statistical maximum of 0.59 +/- 0.1 microm after only 5 minutes of exposure to 500 or 1,000 nM HETE. Exposure of the cornea to 15-(S)-HETE causes a rapid-onset increase in the thickness of a layer of mucin-like glycoprotein on the surface of the corneal epithelium. This supports previous reports that corneal epithelial cells produce mucin and suggests that treatment with topical 15-(S)-HETE may be effective in treating ocular surface mucin deficiency in dry eye syndrome.

Potential for Hospital Based Corneal Retreival in Hassan District Hospital

PubMed Central

Melsakkare, Suresh Ramappa; Manipur, Sahana R.; Acharya, Pavana; Ramamurthy, Lakshmi Bomalapura

2015-01-01

Context In developing countries, corneal diseases are the second leading cause of blindness. This corneal blindness can be treated through corneal transplantation. Though the present infrastructure is strong enough to increase keratoplasty numbers at a required rate, India has largest corneal blind population in the world. So a constant supply of high quality donor corneal tissue is the key factor for reduction of prevalence of corneal blindness. Considering the magnitude of corneal blindness and shortage of donor cornea, there is a huge gap in the demand and supply. Aim To study the potential for hospital based retrieval of donor corneal tissue in Hassan district hospital after analysing the indicated and contraindicated causes of deaths, so that hospital corneal retrieval program in Hassan district hospital can be planned. Materials and Methods The cross-sectional, retrospective and record-based study included all hospital deaths with age group more than two years occurred during one year period (January 2014 to December 2014). Data regarding demographic profile, cause of death, treatment given and presence of any systemic diseases were collected. The causes of deaths which are contraindicated for the retrieval of corneas were analysed and noted. The contraindications were based on the NPCB guidelines for standard of eye banking in India 2009. Results Out of 855 deaths, number of deaths in males (565) was greater than females (290). Numbers of deaths were highest between 41-60 years age group (343). Deaths due to HIV, septicaemia, meningitis, encephalitis, disseminated malignancies were contraindicated for corneal retrieval. Corneas could be retrieved from 736 deaths out of 855. Potential for corneal retrieval in a period of one year in Hassan District hospital was 86%. Conclusion Hospital corneal retrieval program has got a great potential to bridge the gap between the need for the cornea and actually collected corneas which will contribute enormously in eliminating corneal blindness. In present study there was 86% potential for corneal retrieval among the hospital deaths. PMID:26435976

Intra-tissue Refractive Index Shaping (IRIS) of the cornea and lens using a low-pulse-energy femtosecond laser oscillator

PubMed Central

Ding, Li; Knox, Wayne H.; Bühren, Jens; Nagy, Lana J.; Huxlin, Krystel R.

2009-01-01

Purpose To assess the optical effect of high-repetition-rate, low energy femtosecond laser pulses on lightly-fixed corneas and lenses. Methods Eight corneas and eight lenses were extracted post-mortem from normal, adult cats. They were lightly fixed and stored in a solution that minimized swelling and opacification. An 800nm Ti:Sapphire femtosecond laser oscillator with a 27fs pulse duration and 93MHz repetition rate was used to inscribe gratings consisting of 20-40 lines, each 1μm wide, 100μm long and 5μm apart, 100μm below the tissue surface. Refractive index changes in the micromachined regions were calculated immediately and after one month of storage by measuring the intensity distribution of diffracted light when the gratings were irradiated with a 632.8nm He-Ne laser. Results Periodic gratings were created into the stromal layer of the corneas and the cortex of the lenses by adjusting the laser pulse energy until visible plasma luminescence and bubbles were no longer generated. The gratings had low scattering loss and could only be visualized using phase microscopy. Refractive index changes measured 0.005±0.001 to 0.01±0.001 in corneal tissue and 0.015±0.001 to 0.021±0.001 in the lenses. The gratings and refractive index changes were preserved after storing the micromachined corneas and lenses for one month. Conclusions These pilot experiments demonstrate a novel application of low-pulse-energy, MHz femtosecond lasers in modifying the refractive index of transparent ocular tissues without apparent tissue destruction. Although it remains to be verified in living tissues, the stability of this effect suggests that the observed modifications are due to long-term molecular and/or structural changes. PMID:18641284

Comparison of iCare tonometer and Goldmann applanation tonometry in normal corneas and in eyes with automated lamellar and penetrating keratoplasty

PubMed Central

Salvetat, M L; Zeppieri, M; Miani, F; Tosoni, C; Parisi, L; Brusini, P

2011-01-01

Purpose To compare intraocular pressure (IOP) measurements with Goldmann applanation tonometry (GAT) and iCare tonometry in normal and post-keratoplasty corneas and to assess the influence of central corneal thickness (CCT), corneal curvature (CC), and corneal astigmatism (CA) on IOP. Methods This prospective cross-sectional study included one eye of 101 subjects with normal corneas (58 healthy subjects, 43 glaucoma); and 90 post-keratoplasty patients: 34 penetrating keratoplasties (PK); 20 automated-lamellar-therapeutic keratoplasties (ALTK); 19 Descemet-stripping-automated-endothelial keratoplasties (DSAEK); 17 edematous grafts. All subjects underwent GAT and iCare IOP measurements in random order, and CCT, CC, and CA evaluation. The Bland–Altman method and multivariate regression analysis were used to assess inter-tonometer agreement and the influence of CCT, CC, and CA on IOP. Results iCare significantly underestimated IOP in all groups compared with GAT (GAT minus iCare of 3.5±3.5 mm Hg, P<0.001), but overestimated IOP in the edematous grafts (GAT minus iCare of −6.5±1.9 mm Hg, P<0.001). In normal corneas, both tonometer measurements were directly related to CCT values; iCare readings appeared inversely related to CC. There was no significant relationship between IOP and CCT, CC and CA in post-keratoplasty eyes, except between CC and iCare measurements for PK eyes. Conclusions The agreement between GAT and iCare was clinically acceptable in control, ALTK and DSAEK groups, and poor in PK and edematous grafts eyes. In normal corneas, GAT was significantly affected by CCT; iCare was influenced by CCT and CC. The iCare appeared less influenced by corneal edema when compared with GAT. High IOP readings taken with both tonometers in grafts should raise suspicion of true elevated IOP. PMID:21436848

Cornea Optical Topographical Scan System (COTSS)

NASA Technical Reports Server (NTRS)

1986-01-01

The Cornea Optical Topographical Scan System (COTSS) is an instrument designed for use by opthalmologist to aid in performing surgical procedures such as radial keratotomy and to provide quick accurate data to aid in prescribing contact lenses and eyeglasses. A breadboard of the system was built and demonstrated in June of 1984. Additional refinements to the breadboard are needed to meet systems requirements prior to proceeding with prototype development. The present status of the COTSS instrument is given and the areas in which system refinements are required, are defined.

13TH Scientific Session of the Ukrainian Scientific Research Institute of Eye Diseases Imeni Porf. L. L. Girshman.

DTIC Science & Technology

1960-07-29

changes. A broad application of ascorbic acid in the therapy of inflammatory and degenerative ailments of the cornea was proposed. Ye. M. Orlova...accumulation of cholesterol and phosphatides occurs, leading to an opacity of the fibers. In X-ray cataract the opacity of the fibers apparently occurs...corneal spots and the indices he had developed for repeated transplant of the cornea . The reports of Cand. of Med. Sei. N. I. Pil’man (Kiev; and Cand

[Costs of preserved corneal transplants].

PubMed

Ardjomand, N; Reich, M E

1997-10-01

Organ culture medium and Optisol are the most commonly used corneal storage mediums. This study compares the costs for these two methods. In the calculation of costs we did not just take the direct costs into account, but also tried to determine the fixed costs per transplanted cornea with corresponding assumptions. Proceeding on the assumption that 50 stored corneas were transplanted per year, an amount of 11,660 ATS (1,666 DM, 857 ECU) for each organ cultured and 11,986 ATS (1,712 DM, 881 ECU) for each graft preserved in Optisol was calculated. Raising the number of transplanted corneas to 400 per year, each tissue stored in organ culture medium costs 2,811 ATS (402 DM, 207 ECU) and those preserved in Optisol 3234 ATS (462 DM, 238 ECU). Since organ culture storage gives us a reduction in costs of more than 15% compared to storing in Optisol, when preserving 400 transplantable grafts, from the business economics aspect, this storage method should be preferred.

Physical Aspects of Photodynamic Corneal Collagen Crosslinking

NASA Astrophysics Data System (ADS)

Kornfield, Julia

2012-02-01

Healthy vision depends on the stability of the shape of the cornea, which provides most of the lens power of the optical system of the eye. Diseases in which the cornea progressively undergoes irregular deformation over time (e.g., keratoconus) can be treated clinically by inducing additional protein-protein crosslinks using a photosensitizing drug and a tailored dose of light. Unfortunately, the treatment moving through clinical trials is toxic to cells in and on the cornea. A path to a safer treatment is offered by the nanostructure of the corneal stroma---reminiscent of a HEX phase in block copolymers with 30nm diameter collagen cylinders spaced 60nm center-to-center in a hydrogel matrix of proteoglycans and water. We show that using a photosensitizing drug that sequesters itself in the collagen fibrils can minimize the toxicity of therapeutic protein-protein cross-linking. Photorheology and transport measurements are used to quantify the parameters of a simple physical model that is useful for optimizing clinical protocols.

Chemical Sympathectomy Increases Susceptibility to Ocular Herpes Simplex Virus Type 1 Infection

PubMed Central

Templeton, Amanda; Nguyen, Gabrielle; Ash, John D.; Straub, Rainer H.; Carr, Daniel J. J.

2008-01-01

The cornea is one of the most highly innervated tissues in the mammalian host. We hypothesized changes to cornea innervation through chemical sympathectomy would significantly alter the host response to the neurotropic viral pathogen, herpes simplex virus type 1 (HSV-1) following ocular infection. Mice treated with 6-hydroxydopamine hydrobromide displayed reduced tyrosine hydroxylase-positive fibers residing in the cornea. Sympathectomized mice were also found to show a transient rise in virus recovered in infected tissues and succumbed to infection in greater numbers. Whereas there were no differences in infiltrating leukocyte populations including HSV-1-specific cytotoxic T lymphocytes in the infected tissue, an increase in substance P and a decrease in IFN-γ levels in the trigeminal ganglion but not brain stem of sympathectomized mice were noted. Sympathectomized mice treated with the neurokinin-1 receptor antagonist L703,606 had delayed mortality implicating the involvement of substance P in HSV-1-mediated death. PMID:18495255

Corneal birefringence measured by spectrally resolved Mueller matrix ellipsometry and implications for non-invasive glucose monitoring

PubMed Central

Westphal, Peter; Kaltenbach, Johannes-Maria; Wicker, Kai

2016-01-01

A good understanding of the corneal birefringence properties is essential for polarimetric glucose monitoring in the aqueous humor of the eye. Therefore, we have measured complete 16-element Mueller matrices of single-pass transitions through nine porcine corneas in-vitro, spectrally resolved in the range 300…1000 nm. These ellipsometric measurements have been performed at several angles of incidence at the apex and partially at the periphery of the corneas. The Mueller matrices have been decomposed into linear birefringence, circular birefringence (i.e. optical rotation), depolarization, and diattenuation. We found considerable circular birefringence, strongly increasing with decreasing wavelength, for most corneas. Furthermore, the decomposition revealed significant dependence of the linear retardance (in nm) on the wavelength below 500 nm. These findings suggest that uniaxial and biaxial crystals are insufficient models for a general description of the corneal birefringence, especially in the blue and in the UV spectral range. The implications on spectral-polarimetric approaches for glucose monitoring in the eye (for diabetics) are discussed. PMID:27446644

Cross-linking and the molecular packing of corneal collagen

NASA Technical Reports Server (NTRS)

Yamauchi, M.; Chandler, G. S.; Tanzawa, H.; Katz, E. P.

1996-01-01

We have quantitatively characterized, for the first time, the cross-linking in bovine cornea collagen as a function of age. The major iminium reducible cross-links were dehydro-hydroxylysinonorleucine (deH-HLNL) and dehydro-histidinohydroxymerodesmosine (deH-HHMD). The former rapidly diminished after birth; however, the latter persisted in mature animals at a level of 0.3 - 0.4 moles/mole of collagen. A nonreducible cross-link, histidinohydroxylysinonorleucine (HHL), previously found only in skin, was also found to be a major mature cross-link in cornea. The presence of HHL indicates that cornea fibrils have a molecular packing similar to skin collagen. However, like deH-HHMD, the HHL content in corneal fibrils only reaches a maximum value with time about half that of skin. These data suggest that the corneal fibrils are comprised of discrete filaments that are internally stabilized by HHL and deH-HHMD cross-links. This pattern of intermolecular cross-linking would facilitate the special collagen swelling property required for corneal transparency.

Staphylococcus aureus causes acute inflammatory episodes in the cornea during contact lens wear.

PubMed

Wu, P Z; Thakur, A; Stapleton, F; Willcox, M D

2000-06-01

The aim of this investigation was to determine the ability of Staphylococcus aureus to cause keratitis during contact lens wear in the rabbit. Rabbits were fitted with hydrogel lenses and wore them on an extended wear schedule for 7 weeks. At the end of each week of wear, one drop of S. aureus was added to the eyes, and the eyes were monitored using slit lamp bio-microscopy, with subsequent microbial and histological studies of the cornea. In the presence of S. aureus, keratitis occurred after 3 weeks of lens wear, a condition that was characterized by diffuse infiltration of the peripheral cornea and limbal redness. In the absence of bacteria, there was no infiltration until week 6. Contact lens wear also prolonged the retention of bacteria in the eye. Extended contact lens wear and S. aureus caused non-ulcerative keratitis in a rabbit model, and disturbances in the ability of the eye to remove bacteria.

[What should we think? Is pneumotonometry still reliable?].

PubMed

Bresson-Dumont, H; Lehoux, A; Kponoume, C; Santiago, P-Y

2007-05-01

Noncontact tonometer (NCT) is an applanation tonometry, as is Goldmann tonometry. Numerous studies have shown the reliability of this technique but problems still exist. We measured intraocular pressure (IOP) in 340 patients (668 eyes) with NCT and Goldmann tonometer (GAT). To evaluate the influence of central corneal thickness (CCT), we classified the patients according to their CCT (<500 microm; between 520 and 580 microm, and>600 microm). The mean difference between the two IOP measurements was -1.4 mmHg +/- 2.67 (-12 to +7), p<0,0001 for the thinner corneas, +0.13 mmHg +/- 2.67 (-8 to +8), p=0.445 for the regular corneas, and + 3.04 mmHg +/- 3.2 (-7 to +13) p<0.0001 for the thicker corneas. CCT influences NCT and GAT, but the difference between the two techniques is higher for thicker CCTs, higher IOPs, and in younger patients. NCT is good for glaucoma screening but is less reliable than GAT for glaucoma follow-up if CCT and IOP are far from regular values.

Pre- and postoperative optical resolution of the cornea: a preliminary report

NASA Astrophysics Data System (ADS)

Parel, Jean-Marie A.; Simon, Gabriel; Rol, Pascal O.; Ren, Qiushi; Lee, William E.

1993-06-01

The effect of novel refractive surgical techniques on visual acuity and contrast sensitivity is normally determined by the outcome of human clinical trials. For example, ArF laser photorefractive keratoplasty follows an algorithm based on the patient's preoperative data for keratometry, refraction, pachometry, and ocular length all measured with ultrasound. A normalized ablation rate (which is function of the laser fluence), and the desired refractive correction are then used to calculate the ablation depth. On the day of surgery, the epithelium is mechanically removed and the bare cornea photoablated. Finally, the cornea may be medicated with a topical application of antibiotics and the eye is patched. On postoperative day 7, the epithelium is healed and visual acuity and keratometry are measured. With PRK, the theoretical outcome refraction should be within +/- 0.25 D. Thus far however, reproducibility is only of +/- 2 D. We believe the large discrepancy between theory and practice is due to several parameters that vary patient-to-patient.

The G-protein-coupled receptor, GPR84, is important for eye development in Xenopus laevis.

PubMed

Perry, Kimberly J; Johnson, Verity R; Malloch, Erica L; Fukui, Lisa; Wever, Jason; Thomas, Alvin G; Hamilton, Paul W; Henry, Jonathan J

2010-11-01

G-protein-coupled receptors (GPCRs) represent diverse, multifamily groups of cell signaling receptors involved in many cellular processes. We identified Xenopus laevis GPR84 as a member of the A18 subfamily of GPCRs. During development, GPR84 is detected in the embryonic lens placode, differentiating lens fiber cells, retina, and cornea. Anti-sense morpholino oligonucleotide-mediated knockdown and RNA rescue experiments demonstrate GPR84's importance in lens, cornea, and retinal development. Examination of cell proliferation using an antibody against histone H3 S10P reveals significant increases in the lens and retina following GPR84 knockdown. Additionally, there was also an increase in apoptosis in the retina and lens, as revealed by TUNEL assay. Reciprocal transplantation of the presumptive lens ectoderm between uninjected controls and morpholino-injected embryos demonstrates that GPR84 is necessary in the retina for proper development of the retina, as well as other eye tissues including the lens and cornea. © 2010 Wiley-Liss, Inc.

The G-protein-coupled receptor, GPR84, is important for eye development in Xenopus laevis

PubMed Central

Perry, Kimberly J.; Johnson, Verity R.; Malloch, Erica L.; Fukui, Lisa; Wever, Jason; Thomas, Alvin G.; Hamilton, Paul W.; Henry, Jonathan J.

2010-01-01

G-protein-coupled receptors (GPCRs) represent diverse, multifamily groups of cell signaling receptors involved in many cellular processes. We identified Xenopus laevis GPR84 as a member of the A18 subfamily of GPCRs. During development, GPR84 is detected in the embryonic lens placode, differentiating lens fiber cells, retina and cornea. Anti-sense morpholino oligonucleotide-mediated knockdown and RNA rescue experiments demonstrate GPR84’s importance in lens, cornea and retinal development. Examination of cell proliferation using an antibody against histone H3 S10P reveals significant increases in the lens and retina following GPR84 knockdown. Additionally, there was also an increase in apoptosis in the retina and lens, as revealed by TUNEL assay. Reciprocal transplantation of the presumptive lens ectoderm between uninjected controls and morpholino injected embryos demonstrates that GPR84 is necessary in the retina for proper development of the retina, as well as other eye tissues including the lens and cornea. PMID:20925114

Combined application of prophylactic corneal cross-linking and laser in-situ keratomileusis - a review of literature.

PubMed

Chan, Tommy C Y; Ng, Alex L K; Chan, Karen K W; Cheng, George P M; Wong, Ian Y H; Jhanji, Vishal

2017-11-01

Laser in-situ keratomileusis (LASIK) is safe and effective laser refractive procedures in treating refractive errors. However, regression of treatment and iatrogenic keratectasia remain to be a major concern, especially in treating thin cornea with high ametropia. Collagen cross-linking (CXL) is an effective method in stopping keratoconus progression through increasing the biomechanical strength of the cornea. Adjuvant cross-linking to refractive procedures can theoretically help prevent regression and reduce the risk of keratectasia development by increasing the mechanical stability of cornea. During the procedure, riboflavin is directly applied to the corneal stroma, thereby reducing the need of de-epithelialization as in the conventional protocol for keratoconus. Currently, there is still no consensus regarding the indication of CXL during refractive procedure, nor any standardized treatment protocol. This article aims to summarize the current evidence regarding the use of adjuvant CXL in LASIK. © 2016 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Imaging shear stress distribution and evaluating the stress concentration factor of the human eye

NASA Astrophysics Data System (ADS)

Joseph Antony, S.

2015-03-01

Healthy eyes are vital for a better quality of human life. Historically, for man-made materials, scientists and engineers use stress concentration factors to characterise the effects of structural non-homogeneities on their mechanical strength. However, such information is scarce for the human eye. Here we present the shear stress distribution profiles of a healthy human cornea surface in vivo using photo-stress analysis tomography, which is a non-intrusive and non-X-ray based method. The corneal birefringent retardation measured here is comparable to that of previous studies. Using this, we derive eye stress concentration factors and the directional alignment of major principal stress on the surface of the cornea. Similar to thermometers being used for monitoring the general health in humans, this report provides a foundation to characterise the shear stress carrying capacity of the cornea, and a potential bench mark for validating theoretical modelling of stresses in the human eye in future.

In vivo laser confocal microscopic analysis of corneal K-structures after keratorefractive surgery (LASIK and epi-LASIK).

PubMed

Yokogawa, Hideaki; Kobayashi, Akira; Tagawa, Kosaku; Sugiyama, Kazuhisa

2010-01-01

To demonstrate alterations of corneal K-structures (sub-Bowman's fibrous structures) after keratorefractive surgery by in vivo laser confocal microscopy and to look for association of K-structures with fluorescein-stained anterior corneal mosaic (ACM). Five patients (nine eyes) participated in this study. For four patients, one eye was evaluated after laser in situ keratomileusis (LASIK) and the other after epipolis-laser in situ keratomileusis (epi-LASIK). For one patient, the left eye was evaluated after epithelial debridement. A photograph of the ACM was obtained. Central corneal regions were scanned by Heidelberg Retina Tomograph 2 Rostock Cornea Module (Heidelberg Engineering GmbH, Heidelberg, Germany). The ACM and K-structures disappeared in all corneas after epi-LASIK, but not after LASIK and epithelial debridement cornea. The presence of K-structures and ACM may be an index to identify eyes that had a previous refractive surgical procedure (surface ablation or LASIK) and be a health index of Bowman layer and adjacent anterior stroma. Copyright 2010, SLACK Incorporated.

[Evaluation of Iris Morphology Viewed through Stromal Edematous Corneas by Infrared Camera].

PubMed

Kobayashi, Masaaki; Morishige, Naoyuki; Morita, Yukiko; Yamada, Naoyuki; Kobayashi, Motomi; Sonoda, Koh-Hei

2016-02-01

We reported that the application of infrared camera enables us to observe iris morphology in Peters' anomaly through edematous corneas. To observe the iris morphology in bullous keratopathy or failure grafts with an infrared camera. Eleven bullous keratopathy or failure grafts subjects (6 men and 5 women, mean age ± SD; 72.7 ± 13.0 years old) were enrolled in this study. The iris morphology was observed by applying visible light mode and near infrared light mode of infrared camera (MeibomPen). The detectability of pupil shapes, iris patterns and presence of iridectomy was evaluated. Infrared mode observation enabled us to detect the pupil shapes in 11 out of 11 cases, iris patterns in 3 out of 11 cases, and presence of iridetomy in 9 out of 11 cases although visible light mode observation could not detect any iris morphological changes. Applying infrared optics was valuable for observation of the iris morphology through stromal edematous corneas.

Topical timolol with and without benzalkonium chloride: epithelial permeability and autofluorescence of the cornea in glaucoma.

PubMed

de Jong, C; Stolwijk, T; Kuppens, E; de Keizer, R; van Best, J

1994-04-01

Epithelial permeability and autofluorescence of the cornea were determined by fluorophotometry in 21 patients with open-angle glaucoma or ocular hypertension using timolol medication with the preservative benzalkonium chloride (BAC) and 2 weeks after changing to timolol medication without BAC. The investigation was performed to determine whether removal of BAC would reduce toxic effects on the cornea and complaints of sensations of burning or dry eye. Corneal epithelial permeability decreased significantly after changing medication (mean decrease per patient 27%, P = 0.025). Corneal autofluorescence increased significantly after changing medication suggesting an alteration in corneal metabolism (mean increase per patient 6%, P = 0.003). Timolol without BAC was found to be as effective as timolol with BAC in reducing intraocular pressure (P = 0.4). Removal of BAC from timolol resulted in an improvement of corneal epithelial barrier function and in a reduction of complaints. The improvement was found to be proportional to the duration of the preceding BAC-containing therapy.

Corneal injury to ex vivo eyes exposed to a 3.8-micron laser

NASA Astrophysics Data System (ADS)

Fyffe, James G.; Randolph, Donald Q.; Winston, Golda C. H.; Johnson, Thomas E.

2005-04-01

As a consequence of the enormous expansion of laser use in medicine, industry and research, specific safety standards must be developed that appropriately address eye protection. The purpose of this study is to establish injury thresholds to the cornea for 3.8 micron 8 microsecond laser light pulses and to investigate a possible replacement model to live animal testing. Previous studies of pulsed energy absorption at 3.8 microns were performed using rhesus monkey cornea and were at pulse durations two orders of magnitude different than the 8 microsecond pulses used in this study. Ex-vivo pig eyes were exposed at varying energies and evaluated to establish the statistical threshold for corneal damage. Histology was used to determine the extent of damage to the cornea. It is expected that the results will be used to assist in the establishment of safety standards for laser use and offer an alternative to future animal use in establishment of safety standards.

Femtosecond laser corneal surgery with in situ determination of the laser attenuation and ablation threshold by second harmonic generation

NASA Astrophysics Data System (ADS)

Plamann, Karsten; Nuzzo, Valeria; Albert, Olivier; Mourou, Gérard A.; Savoldelli, Michèle; Dagonet, Françoise; Donate, David; Legeais, Jean-Marc

2007-02-01

Femtosecond lasers start to be routinely used in refractive eye surgery. Current research focuses on their application to glaucoma and cataract surgery as well as cornea transplant procedures. To avoid unwanted tissue damage during the surgical intervention it is of utmost importance to maintain a working energy just above the ablation threshold and maintain the laser energy at this working point independently of the local and global tissue properties. To quantify the attenuation of the laser power density in the tissue by absorption, scattering and modification of the point spread function we monitor the second harmonic radiation generated in the collagen matrix of the cornea when exposed to ultrashort laser pulses. We use a CPA system with a regenerative amplifier delivering pulses at a wavelength of 1.06 μm, pulse durations of 400 fs and a maximum energy of 60 μJ. The repetition rate is adjustable from single shot up to 10 kHz. The experiments are performed on human corneas provided by the French Eye bank. To capture the SHG radiation we use a photomultiplier tube connected to a lockin amplifier tuned to the laser repetition rate. The measured data indicates an exponential decay of the laser beam intensity in the volume of the sample and allows for the quantification of the attenuation coefficient and its correlation with the optical properties of the cornea. Complementary analyses were performed on the samples by ultrastructural histology.

Cellular and nerve regeneration within a biosynthetic extracellular matrix for corneal transplantation

NASA Astrophysics Data System (ADS)

Li, Fengfu; Carlsson, David; Lohmann, Chris; Suuronen, Erik; Vascotto, Sandy; Kobuch, Karin; Sheardown, Heather; Munger, Rejean; Nakamura, Masatsugu; Griffith, May

2003-12-01

Our objective was to determine whether key properties of extracellular matrix (ECM) macromolecules can be replicated within tissue-engineered biosynthetic matrices to influence cellular properties and behavior. To achieve this, hydrated collagen and N-isopropylacrylamide copolymer-based ECMs were fabricated and tested on a corneal model. The structural and immunological simplicity of the cornea and importance of its extensive innervation for optimal functioning makes it an ideal test model. In addition, corneal failure is a clinically significant problem. Matrices were therefore designed to have the optical clarity and the proper dimensions, curvature, and biomechanical properties for use as corneal tissue replacements in transplantation. In vitro studies demonstrated that grafting of the laminin adhesion pentapeptide motif, YIGSR, to the hydrogels promoted epithelial stratification and neurite in-growth. Implants into pigs' corneas demonstrated successful in vivo regeneration of host corneal epithelium, stroma, and nerves. In particular, functional nerves were observed to rapidly regenerate in implants. By comparison, nerve regeneration in allograft controls was too slow to be observed during the experimental period, consistent with the behavior of human cornea transplants. Other corneal substitutes have been produced and tested, but here we report an implantable matrix that performs as a physiologically functional tissue substitute and not simply as a prosthetic device. These biosynthetic ECM replacements should have applicability to many areas of tissue engineering and regenerative medicine, especially where nerve function is required. regenerative medicine | tissue engineering | cornea | implantation | innervation

[Expression of matrix metalloproteinase-19 in the human cornea. Wound healing in the MMP-19 knock-out mouse model].

PubMed

Treumer, F; Flöhr, C; Klettner, A; Nölle, B; Roider, J

2010-07-01

At present there are no data in the literature on the expression of matrix metalloprotein-19 in the human cornea. The aim of this study was to analyze the expression of matrix metalloproteinase-19 in the human cornea and to investigate its potential role in corneal wound healing using a MMP-19 knock-out mouse model. A method with Western blotting and immunohistological staining for MMP-19 was performed using paraffin embedded human corneas. Excimer laser keratectomy was performed in wild type (wt) and MMP-19 knock-out (ko) mice and the rate of re-epithelialization was analyzed after 8 h and 18 h. MMP-19 was strongly expressed in the human corneal epithelium mainly in the basal cell layer. MMP-19 was not expressed in the corneal stroma. In the mouse model the size of the corneal lesion after 8 h was 83% (wt) and 89.9% (ko) of the initial area (p=0.09). After 18 h the lesion was 17% (wt) and 13.3% (ko) of the initial area (p=0.01). Laminin-5 was expressed in the migrating epithelial cells with no differences between wild type and knock-out mouse. MMP-19 showed a strong expression in the basal cells of the human corneal epithelium. Corneal re-epithelialization was slightly faster in the MMP-19 knock-out mouse. No differences in the expression of laminin-5 could be detected.

Impact of Hydration Media on Ex Vivo Corneal Elasticity Measurements

PubMed Central

Dias, Janice; Ziebarth, Noël M.

2014-01-01

Objectives To determine the effect of hydration media on ex vivo corneal elasticity. Methods Experiments were conducted on forty porcine eyes retrieved from an abattoir (10 eyes each for PBS, BSS, Optisol, 15% Dextran). The epithelium was removed and the cornea was excised with an intact scleral rim and placed in 20% Dextran overnight to restore its physiological thickness. For each hydration media, corneas were evenly divided into two groups: one with an intact scleral rim and the other without. Corneas were mounted onto a custom chamber and immersed in a hydration medium for elasticity testing. While in each medium, corneal elasticity measurements were performed for 2 hours: at 5-minute intervals for the first 30 minutes and then 15-minute intervals for the remaining 90 minutes. Elasticity testing was performed using nanoindentation with spherical indenters and Young’s modulus was calculated using the Hertz model. Thickness measurements were taken before and after elasticity testing. Results The percentage change in corneal thickness and elasticity was calculated for each hydration media group. BSS, PBS, and Optisol showed an increase in thickness and Young’s moduli for corneas with and without an intact scleral rim. 15% Dextran exhibited a dehydrating effect on corneal thickness and provided stable maintenance of corneal elasticity for both groups. Conclusions Hydration media affects the stability of corneal thickness and elasticity measurements over time. 15% Dextran was most effective in maintaining corneal hydration and elasticity, followed by Optisol. PMID:25603443