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Purpose: To characterize and correlate guttata severity, Descemet's membrane thickness (DMT), central cornea thickness (CCT) in corneas with guttae using specular microscopy and spectral-domain optical coherence tomography (SD-OCT) and test the Doheny Image Reading Center (DIRC) specular microscopy-based corneal guttata severity scale. Methods: Forty-nine eyes of 49 patients with guttata and 36 eyes of age-matched of 36 normal controls were enrolled in the study. Three images of the central cornea and four of the peripheral cornea (inferior, superior, nasal and temporal) of each eye were taken using the Konan NSP-9900 specular microscope. A volume scan of the central cornea cross-section was collected on each eye using the Heidelberg Spectralis SD-OCT. The density of endothelial guttata based on specular images was graded on a 0–4 scale, and the Descemet's membrane thickness (DMT) and central corneal thickness (CCT) were manually measured by two trained graders. Results: The DIRC corneal guttata severity scale showed good reproducibility of all corneal endothelial images (weighted Kappa = 0.87). Mean DMT was 16.1 ± 2.4 µm in controls and 25.5 ± 10.9 µm in corneas with guttata (P < 0.001). Mean CCT was 552 ± 26 µm in controls and 603 ± 55 µm in corneas with guttata (P < 0.001). Guttata severity was significantly correlated with both DMT (r = 0.743, P < 0.001) and CCT (r = 0.569, P < 0.001). Age was moderately correlated with DMT (r = 0.472, P = 0.003) and mildly correlated with guttata severity (r = 0.285, P = 0.031), but was not correlated with CCT (r = 0.058, P = 0.681). Guttatta grade 3 corneas displayed an increase in DMT and guttata grade 4 was associated with a significant increase in CCT. Conclusions: The DMT and CCT are increased in corneas with guttata. The higher density of guttae is correlated with increased thickness. Specular microscopy combined with SD-OCT can be used as a good approach to assess the severity of FECD. [ABSTRACT FROM AUTHOR]

Purpose: To measure variation in corneal dendritic cell density, and percentage of mature to total dendritic cells, in healthy individuals during the sleep/wake cycle. Methods: Using in vivo confocal microscopy, images of the subbasal nerve plexus were captured from 19 healthy, noncontact lens wearing participants. The central cornea and inferior whorl were imaged three times (midday, before sleep, upon awakening). Dendritic cell counts from the images were categorized according to perceived maturity (immature vs mature). Dendritic cell density and percentage of mature to total cells were compared between time points. Result: The median and interquartile range (IQR) of total dendritic cell density in the central cornea was 32.0 (7.0–131.3) cells/mm2 at midday, 37.1 (8.2–103.9) cells/mm2 before sleep, and 19.5 (7.0–83.2) cells/mm2 on awakening. Corresponding values for immature cells were 28.1 (5.8–112.5) cells/mm2, 22.3 (7.4–84.0) cells/mm2 and 18.0 (2.9–64.8) cells/mm2, and for mature cells, 3.1 (0.0–6.6) cells/mm2, 2.0 (0.8–16.8) cells/mm2, and 1.6 (0.2–8.2) cells/mm2. At the inferior whorl, total dendritic cell density was 38.5 (18.4–84.5) cells/mm2, 34.4 (9.4–82.3) cell/mm2, and 32.3 (15.2–96.1) cells/mm2. Immature cell density was 32.8 (18.4–80.9) cells/mm2, 34.4 (8.6–81.0) cells/mm2, and 32.3 (12.6–78.5) cells/mm2. Mature cell density was 1.6 (0.0–6.3) cells/mm2, 1.6 (0.0–3.1) cells/mm2, and 1.8 (0.0–6.3) cells/mm2. There was no significant difference between time points for total cell density (p > 0.05), but the percentage of mature cells upon awakening was significantly greater, compared to midday, at the central cornea (p = 0.02). Conclusion: In healthy individuals, overall corneal dendritic cell density is reasonably constant during the sleep/wake cycle, but the relative number of mature cells tends to increase overnight. [ABSTRACT FROM AUTHOR]

To investigate the effect of different pH phosphate buffer solution (PBS) drops on intact corneal biomechanics. Intact rabbit cornea with a 3 mm scleral skirt was sampled and immediately applied in inflation tests within 5 min. After preconditioning, a stable loading cycle from 0.3 to 6 kPa was performed followed by a 10 min interval. During the interval, the samples were randomized into four groups, with one control group receiving no drops and three groups with PBS drops of pH 6.9, 7.4 or 7.9 administration on the surface once per minute. The pressure and displacement were collected at the baseline and 10, 20 and 30 min following the administration. Continuous corneal thickness increase was manifested following the administration of PBS but not in the control group. There was significant swelling-independent corneal modulus reduction after PBS administration, which occurred mainly in the first 10 min. PBS of pH 6.9 achieved significantly smaller modulus reduction than that with pH 7.4 PBS adjusted for thickness changes (p < 0.05). Pressure-modulus curve linear fitting demonstrated that the curve coefficient significantly reduced following PBS administration, and the coefficient decline was smallest with pH 6.9 PBS among three PBS administration groups (p < 0.05). The study demonstrated that the PBS drops of various pH administration could decrease the corneal stiffness independent of corneal swelling. Following the PBS administration, the stiffness changes were more prominent as the posterior pressure increased, and the minimal effect was achieved with slightly acidic PBS. The research provides the essence for stabilizing the corneal biomechanical properties by regulating the pH value of tear film and intraocular pressure. [ABSTRACT FROM AUTHOR]

Purpose: Corneal neurotization (CN) is a novel, potentially curative surgical procedure for the treatment of neurothophic keratopathy (NK). Patients with severe NK can present with corneal opacification requiring optical keratoplasty, which would likely fail without a proper trophic support of corneal nerves in the recipient cornea. Methods: This is a pilot study on 4 patients undergoing keratoplasty after CN. Pre- and postoperative data at 12, 24 months and at the last follow-up were collected for the examination of (i) best corrected visual acuity (BCVA), (ii) slit lamp examination and photograph acquisition with and without fluorescein staining, (iii) corneal aesthesiometry, (iv) in vivo confocal microscopy of the central cornea. Neurophysiological study of the corneal reflex before corneal graft and at last follow up was performed. Results: Four female patients (47.25 ± 5.06 y.o.) underwent keratoplasty after CN (3 penetrating keratoplasty, 1 deep anterior lamellar keratoplasty). The mean interval between CN and keratoplasty was 22 (± 12) months. The mean graft survival time was 42 (± 25) months. Graft follow-up ranged from 72 to 132 months. At the final follow-up, BCVA was improved in 2 out of 4 patients. The mean corneal sensitivity was 11.9 ± 8.3 mm at last follow-up. In vivo confocal microscopy confirmed the presence of functioning nerves at the last follow-up in all patients. NK-related complications occurred in 3 eyes (2 persistent epithelial defect, 1 corneal melting). The former complication was successfully treated by autologous serum eye drops while the latter required repeated keratoplasty. Conclusions: Keratoplasty is a viable strategy to improve visual acuity in patients with corneal opacity who underwent CN for the treatment of NK. Even in the presence of functioning corneal nerves before keratoplasty, surgeons should be aware of the increased rate of NK-related complications that could require the need for repeated procedure. [ABSTRACT FROM AUTHOR]

Purpose: To compare the refractive efficacy and morphological changes in the cornea following a novel biphasic higher fluence transepithelial corneal crosslinking (BI-TE-CXL) and transepithelial corneal crosslinking (TE-CXL) in adults keratoconus. Methods: Patients with progressive keratoconus who required corneal crosslinking were assigned to the BI-TE-CXL group (32 eyes, phase 1: 7.2 J/cm2 for 5 min and 20 s of pulsed-light exposure, KXL, Glaukos-Avedro; phase 2: 3.6 J/cm2 for 6 min and 40 s of continuous light exposure at the front curvature apex with a 6 mm diameter light spot, UVX-2000, IROC) or the TE-CXL group (32 eyes, uniform 7.2 J/cm2 for 5 min and 20 s of pulsed-light exposure, KXL, Glaukos-Avedro). Uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), corneal fluorescein staining (CFS), corneal topography, anterior segment optical coherence tomography (AS-OCT), and in vivo corneal confocal microscopy (IVCM) were performed 3, 6, 12 and 24 months after surgery. Results: The CFS scores in the BI-TE-CXL group were significantly higher than those in the TE-CXL group on the first two days after surgery (p < 0.001). The Kmax (at 12 and 24 months) and CDVA (logMAR) were significantly lower in the BI-TE-CXL group than those in the TE-CXL group (p < 0.05). The corneal demarcation line under AS-OCT was visible in 81.3% of patients in the BI-TE-CXL group and 15.6% in the TE-CXL group. The depth of the demarcation line under IVCM was significantly deeper in the BI-TE-CXL group (248.3 ± 25.0 μm) than that of the TE-CXL group (136.5 ± 15.6 μm) in the central cornea (p < 0.001). The cross-linked collagen structures in the central cornea were still present after 12 months in the BI-TE-CXL group. No significant difference in sub-basal nerve density between the two groups (p > 0.05). Conclusions: Following BI-TE-CXL, CDVA was significantly improved, accompanied by deeper demarcation line depth and persistent crosslinked structures in the central corneal stroma. [ABSTRACT FROM AUTHOR]

Purpose: Corneal epithelial defects from trauma or surgery heal as new epithelial cells grow centripetally from the limbus and replenish the epithelium. Corneal wound healing requires cell signalling molecules. However, a topical treatment with these components is not available. Human breast milk (HBM) offers a potential, novel treatment as it contains bioactive molecules important in epithelial cell healing. This study seeks to investigate the potential of HBM in cornea wound healing. Methods: Balb/c mice, 8–12 weeks old, were anesthetized prior to creating a 2 mm central cornea epithelial defect. Mice were randomly assigned to a treatment group: HBM, ophthalmic ointment containing neomycin, polymyxin B, dexamethasone (RxTx), or saline and treated 4x/day for 2 days. Wound area was quantified by fluorescein and ImageJ at 0, 8, 24, and 48 h post wounding and eyes used for histology, RT-qPCR, and ELISA. Results: Wounded corneas treated with HBM demonstrated increased re-epithelialization at 8 h post injury compared to saline treatments. ELISA showed significantly higher Ki67 in HBM treated eyes vs. saline control at 8 h (p = 0.0278). Additionally, immunohistology revealed more Ki67 positive cells in the HBM group compared to saline at 8 h and 24 h (p = 0.0063 8 h; p = 0.0007 24 h). For inflammatory analysis, HBM group IL-1β levels were similar to the saline group, and higher than RxTx treated eyes (p < 0.05). Immunohistochemical staining for CD11b (macrophage marker) revealed HBM-treated eyes had significantly more positive cells vs. saline. RT-qPCR of limbal stem cell markers (LESCs) revealed upregulation of Integrin αV at 8 h with HBM vs. saline. Conclusions: HBM treatment on corneas with debridement of epithelium demonstrated improved healing, cellular proliferation, and upregulation of the LESC gene transcript, integrin αV, after wounding. Future studies could investigate LESC response to different signalling molecules in HBM to better understand the efficacy of this potential therapy. [ABSTRACT FROM AUTHOR]

To evaluate the efficacy of topical vancomycin and povidone iodine (PI) application on methicillin-resistant Staphylococcus aureus (MRSA) keratitis model in rabbits. MRSA keratitis was induced by injecting 0.1 mL MRSA containing 1000 colony-forming units (CFU) into central cornea of right eyes of 24 New Zealand White rabbits. Animals were divided into four groups (n = 6): control (treated with balanced salt solution), 50 mg/mL topical vancomycin, 5% topical PI, and combination; examined before and after treatment, and corneal tissues were harvested for analysis at 9th hour of treatment. Bacterial load was determined as: 7.63 ± 0.82 log10 CFU/g in control group, 6.95 ± 1.66 log10 CFU/g in PI group, 4.67 ± 0.77 log10 CFU/g in combination group, and 4.33 ± 0.71 log10 CFU/g in vancomycin group (p = 0.001). Median of total clinical score increased significantly from 7 [range: 5–8] to 11.5 [range: 11–15] (p = 0.001) in control group, did not change (6 [range: 5–8] to 7 [range: 5–7]; p = 0.695) in vancomycin group, increased significantly from 7 [range: 5–8] to 12.5 [range: 10–14] (p < 0.001) in PI group, increased significantly from 6.5 [range: 5–7] to 8 [range: 7–9] in combination group (p = 0.002). Post-treatment clinical scores for chemosis, conjunctival injection, iritis, hypopyon, epithelial erosion, and corneal infiltrate were significantly lower in vancomycin-treated groups compared to others (p < 0.05). In PI-treated groups, especially scores for chemosis, conjunctival injection, epithelial erosion and corneal infiltrate were significantly higher than vancomycin (p < 0.05). Topical vancomycin significantly inhibited bacterial growth in MRSA keratitis. However, PI was ineffective in controlling this growth; additionally, exerted toxic effect on ocular surface. When vancomycin was combined with PI, no additional increase in efficacy of treatment was detected compared to only vancomycin. [ABSTRACT FROM AUTHOR]

Currently, much attention is paid to measuring the temperature of the ocular surface in various ophthalmic diseases. However, for a comprehensive assessment of heat transfer of the eye, it is advisable to measure both the ocular surface temperature and the heat flux (HF) density. This will expand our knowledge of the physiology of the eye and create new possibilities for diagnosing ocular pathology. The present study aimed to develop a thermoelectric device to study rabbits' ocular surface temperature and HF density. The multichannel thermoelectric device was developed to measure the ocular surface temperature and HF density. This study included ten rabbits (20 eyes). In all animals, the temperature and the HF density were measured on the surface of the central cornea of both eyes. The measurement was repeated after pupils' dilation. The corneal surface temperature of rabbits was 33.1 ± 0.8 °C, and the HF density of the surface of the cornea was 8.3 ± 0.6 mW/cm2. Our results revealed a high degree of interocular symmetry in the surface HF density in healthy rabbits. After pupil dilation, an increase in the HF density on the surface of the rabbit cornea compared with the initial data and control eye was noted. The application of the developed device showed that it is safe and allows for the measurement of the ocular surface temperature and the HF density. We believe that further studies on the ocular surface HF density measurement in various eye diseases will allow us to evaluate the possibilities of this device and technique for diagnostic purposes. [ABSTRACT FROM AUTHOR]

Purpose: The role of corneal epithelial dendritic cells (CEDC), a subtype of antigen presenting cells, in ocular allergy remains largely unknown. This cross-sectional study evaluated the density and morphology of CEDC in participants diagnosed with systemic allergy, to increase our understanding of the role of CEDC in ocular inflammation associated with systemic allergy. Materials and methods: A convenience sample of 50 participants was categorised into allergic and non-allergic groups (31 allergic and 19 non-allergic) based on the results of skin prick test (SPT). Ocular allergy symptoms, clinical ocular surface signs and serum IgE were assessed. In vivo confocal microscopy was performed on the right eye only. The number of CEDC in a 1mm2 region at both the central and mid-peripheral cornea was manually counted. CEDC morphology was graded on a 1 to 3 scale. Results: Ocular surface symptoms, signs (other than eyelid oedema), and serum IgE were significantly higher in the allergic (SPT+) group. CEDC density at the mid-peripheral cornea was significantly lower in the allergic group (p =.003). CEDC morphology grades were significantly higher in allergic participants in the central cornea (p =.02), with the highest grade morphology observed only in allergic participants. No associations were evident between CEDC density or morphology and ocular signs, symptoms or serum IgE. Conclusions: The study showed reduced CEDC density and cells with longer dendrites in allergic participants. The more mature CEDC morphology in the allergic group is suggestive of an inflammatory or immune response. [ABSTRACT FROM AUTHOR]

This study aimed to explore whether corneal cross-linking (CXL) could regress corneal blood vessels (CBV) and corneal lymphatic vessels (CLV) in alkali-burned rabbits. A total of 80 rabbits 2–3 months old weighing 1.5–2.0 kg were randomly divided into four groups: CXL7 group; CTL7 group; CXL14 group; and CTL14 group. Then, 3% sodium pentobarbital 1 ml/kg and tetracaine eye drop 5 g/L were administered before surgery. NaOH 2 mol/L was topically applied to the central cornea to establish the alkali burning model. Then CXL was administered within 2 h in groups CXL7 and CXL14. Corneal opacity and edema, CBV and CLV volume, cluster differentiation 31 (CD31), and lymphatic vessel endothelial receptor 1 (LYVE-1) expression levels were analyzed on days 7 and 14. CXL reduced cornea opacity, CNV, and CLV volumes on day 7 in alkali-burned rabbits. However, CNV and CLV volumes were increased on day 14. CXL also showed down- and upregulation of CD31 and LYVE-1 expression levels on days 7 and 14, respectively. CXL effectively regulated CBV and CLV in alkali-burned rabbits. The transient angioregression and lymphangioregression induced by CXL may be potentially helpful in vascularized high-risk eyes. [ABSTRACT FROM AUTHOR]

Purpose: To determine the association between corneal dendritic cell (DC) density and corneal nerve morphology and tear film inflammatory mediators and neuromediators in healthy individuals. Methods: Flush tears were collected from 21 healthy participants aged 39.7 ± 9.9 years and analyzed for total protein content (TPC), substance P, matrix-metalloproteinase-9 (MMP-9), tissue inhibitor of MMPs-1 (TIMP-1), tumor necrosis factor-a (TNF-α) and interleukin-6 (IL-6). In vivo confocal microscopy was used to assess DC density and corneal nerve morphology. Corneal nerve variables measured were corneal nerve fiber length (CNFL), fiber density (CNFD), branch density (CNBD), fiber total branch density (CTBD), fiber area (CNFA), fiber width (CNFW) and fractal dimension (CNFrac). Results: Participants with DC density over 50 cells/mm2 correlated with CNBD-average (r = 0.7, p = 0.02), CNBD-high (r = 0.6, p = 0.02), CNBD-low (r = 0.6, p = 0.02) CTBD-average (r = 0.7, p = 0.01), CTBD-high (r = 0.6, p = 0.03), CTBD-low (r = 0.7, p = 0.01), CNFA-average (r = 0.7, p = 0.00), CNFA-high (r = 0.7, p = 0.01), CNFA-low (r = 0.8, p < 0.001), CNFrac-SD (r = −0.6, p = 0.04), CNFrac-low (r = 0.6, p = 0.04) and CNFL-low (r = 0.7, p = 0.02). The percentage of MMP-9 correlated with DC density in the entire cohort (r = 0.47, p = 0.03). Conclusions: Corneal nerve measures showed a strong correlation with higher DC density, suggesting that the number of cells maybe be modulated by the corneal nerves in the central cornea. MMP-9 also showed a moderate correlation with DC, supporting an inflammatory role. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the influence of benzalkonium chloride (BAC) on corneal Langerhans cells (LCs) and on the development of dry eye. Methods: A randomized double-blind clinical trial was performed in 20 healthy volunteers. One eye of each subject was treated with a 0.01% BAC solution (and the fellow eye with a placebo solution) three times daily for 12 weeks. The distribution and density of LCs in the central and peripheral corneal epithelium were evaluated by in-vivo confocal laser-scanning microscopy (CLSM). The subjects were monitored for dry eye (subjective discomfort, slit-lamp biomicroscopy, tear film break-up time, Schirmer’s test). Results: In the BAC group, compared with placebo, a marked increase in LC density was found in the central cornea at Week 6 and in the central and peripheral cornea at Week 12. LC density then decreased again in both zones after the end of treatment, falling toward (or even below) baseline levels. Significant changes in the LC count relative to baseline were found at Week 12 in the central and peripheral cornea in the BAC group as well as in the peripheral cornea in the placebo group. LC density in the BAC group increased more rapidly in the central than in the peripheral cornea. During therapy the BAC group showed no signs of dry eye. Conclusion: 12-week application of a 0.01% BAC solution in healthy volunteers induces a significant increase in LCs in the central cornea at Week 12 without dry-eye changes. The return of LC counts toward (or even below) baseline levels just four weeks after the end of BAC administration demonstrates the rapid normalization of the inflammatory environment. [ABSTRACT FROM AUTHOR]

Purpose: To measure the cell size and cell density in five layers of the central cornea in the widely used inbred C57BL/6 mouse strain usingin vivothree-dimensional (3D) two-photon (2PH) imaging. Methods: Corneas were scanned using a 2PH laser scanning fluorescence microscope after staining with plasma membrane stain and Hoechst 33342. Good quality 3D images were selected for the cell density and cell size analysis. Cell density was determined by counting the cell nuclei in a predefined cube of 3D images. Cell size measurements, including cell surface area, cell volume, nuclear surface area and nuclear volume, were automatically quantified using the Imaris software. The cell and nuclear surface-area-to-volume ratio (S:V ratio) and the cell nuclear-cytoplasmic ratio (N:C ratio) were calculated. Results: The highest cell density was observed in the basal epithelium and the lowest in the posterior stroma. The highest cell surface area was found in the anterior stroma, and the highest cell volume was observed in the superficial epithelium. The lowest cell surface area and cell volume were both found in the basal epithelium. The highest S:V ratio was observed in the basal epithelium and the lowest in the superficial epithelium. The highest cell nuclear surface area and volume were both observed in the superficial epithelium and the lowest in the basal epithelium. The highest cell nuclear S:V ratio was observed in the basal epithelium and the lowest in the superficial epithelium. The highest N:C ratio was found in the basal epithelial cells and the lowest in the posterior keratocytes. Conclusions: We are the first to quantify the cell density and size parameters, including cell surface area and volume, cell nuclear surface area and volume, and the S:V ratio, in the five layers of the central cornea. These data provide important cell morphology features for the study of corneal physiology, pathology and disease in mice, particularly in C57BL/6 mice. [ABSTRACT FROM PUBLISHER]

Purpose: The objective is to analyze the antiangiogenic mechanism of suramab, a pharmaceutical compound of bevacizumab and suramin, in a rabbit model of corneal angiogenesis. Material and Methods: Corneal neovascularization was induced in four groups of six New Zealand White rabbits by applying a filter paper disk soaked in 1 M Na (OH) on the central cornea. Group one was treated after injury with intravenous suramab at a dose equivalent to 3 mg/kg of bevacizumab and 10 mg/kg of suramin. Group two was treated with intravenous bevacizumab (5 mg/kg). Group three was treated with 10 mg/kg of suramin while the control group received no treatment. Digital photographs were taken at days 9, 15, 21, and 35. Neovessel formation was quantified giving a 0-4 score to each quadrant according to the centripetal growth of the longest vessel (neovessel index, NVI). Animals were sacrificed at day 35. Corneas were processed for histology, immunohistochemistry, and Western-blot using primary antibodies against P2X2, basic fibroblast growth factor (bFGF), LYVE-1, PECAM-1, and vascular endothelial growth factor-A (VEGF-A). Results: Suramab significantly reduced neovessel growth (mean NVI: 4.2) compared to bevacizumab (8.4), suramin (7.22), and control animals (12.2) at 35 days post-injury (p < 0.01). A lower protein expression of P2X2, bFGF, LYVE-1, PECAM-1, and VEGF-A was found in the cornea of suramab animals than in the other groups of animals. Conclusions: Joint downregulation of bFGF, P2X2, bFGF, and LYVE-1 constitutes a mechanism that induces greater and longer inhibition of corneal angiogenesis. Results might be relevant to ophthalmic care. Ocular administration of suramab is currently being investigated. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the influence of positional deviations on rebound tonometer measurement of intraocular pressure in humans. Materials and methods: Intraocular pressure was measured using the Icare rebound tonometer on the right eyes of 53 subjects in various conditions as follows: first, at a distance of 4, 6, or 8 mm from the center of the cornea with the probe perpendicular to the corneal plane; then, at 2 mm from the limbus in the nasal and temporal regions with the probe perpendicular to the corneal plane or along the visual axis; and lastly, with the angled probe touching the central cornea at angles of 10° or 20°. Results: Bland-Altman plots between the Goldmann applanation tonometer and rebound tonometer at various conditions revealed 95% limits of agreement ranging from ±4.5 to ±5.6 mm Hg. Intraocular pressures measured using the rebound tonometer were significantly lower than those measured using the Goldmann applanation tonometer when the rebound tonometer probe was placed 2 mm from the limbus in the temporal or nasal regions with the probe along the visual axis or when the probe was angled to touch the central cornea at an angle of 10° or 20°. In other positions, the intraocular pressures measured using the rebound tonometer were not significantly different. The rebound tonometer, noncontact tonometer, and Tonopen XL showed good agreement with the Goldmann applanation tonometer for intraocular pressure readings under optimal conditions. Conclusions: The intraocular pressures determined using the rebound tonometer were approximately equal to those obtained using the Goldmann applanation tonometer when the rebound tonometer measurements were made with the probe perpendicular to the corneal plane, irrespective of the location, that is, at the central cornea or 2 mm from the limbus. [ABSTRACT FROM AUTHOR]

Purpose: To test if relaxin deficiency affects ocular structure and function we investigated expression of relaxin (Rln) and RXFP receptors (Rxfp1, Rxfp2), and compared ocular phenotypes in relaxin gene knockout (Rln−/−) and wild type (Rln+/+) mice. Materials and Methods: Rln, Rxfp1 and Rxfp2 mRNA expression was detected in ocular tissues of Rln+/+ mice using RT-PCR. The eyes of 11 Rln−/− and 5 Rln+/+ male mice were investigated. Corneal and retinal thickness was assessed using optical coherence tomography. Intraocular pressure was measured using a rebound tonometer. Retinal, choroidal and sclera morphology and thickness were evaluated histologically. Eyes were collected and fixed for immunofluorescence staining or used for RNA extraction to evaluate mRNA expression using real-time PCR. Results: Rln mRNA was expressed only in the retina, whereas Rxfp1 transcripts were detected in the retina, cornea and sclera/choroid. Rxfp2 was only present in the cornea. None of these genes were expressed in the lacrimal gland, eyelid or lens. Intraocular pressure was higher and central cornea of Rln−/− mice was significantly thicker and had significantly larger endothelial cells and a lower endothelial cell density than Rln+/+ mice. Immunohistochemistry demonstrated no significant difference in AQP3 and AQP5 staining in the cornea or other regions between wildtype and Rln−/− mice. mRNA expression of Aqp4 was significantly higher in Rln−/− than in Rln+/+ corneas, whereas Col1a2, Mmp9, Timp1 and Timp2 were significantly decreased. Expression of Aqp1, Aqp4, Aqp5, Vim and Tjp1 was significantly decreased in Rln−/− compared to Rln+/+ uvea. No significant differences in these genes were detected in the retina. Retinal, choroidal and scleral thicknesses were not different and morphology appeared normal. Conclusion: The findings indicate that loss of Rln affects expression of several genes in the uvea and cornea and results in thicker corneas with altered endothelial cells. Many of the gene changes suggest alterations in extracellular matrix and fluid transport between cells. [ABSTRACT FROM AUTHOR]

Purpose: To observe the morphology of the corneal cells and corneal nerve fibers in patients with type IV or V hereditary sensory and autonomic neuropathy (HSAN) by in vivo confocal microscopy and elucidate the mechanism leading to the loss of corneal sensation in this disease. Methods: In vivo confocal microscopy was performed on the central cornea of the right eye in 3 patients with HSAN (ages 17, 20, and 32 years), and their corneal morphology was compared with that of 3 healthy subjects (ages 28, 30, and 36 years). Corneal sensation was tested with a Cochet-Bonnet esthesiometer. Results: The superficial epithelial cell density was lower in the HSAN patients compared with the healthy subjects (1525, 1225, and 1250/mm2 vs. 2225, 1750, and 2500/mm2), but the basal epithelial cell density of the patients was similar to that of the healthy subjects. Nerve bundles were clearly observed in the sub-basal nerve plexus layer of the cornea in the healthy subjects, but were undetectable at the central cornea in the patients with HSAN. The corneal sensation of the patients with HSAN was much weaker than that of the healthy subjects (2.79, 40.30, and 132.50 g/mm2 vs. 1.47, 1.47, and 1.47 g/mm2). Conclusions: Superficial keratopathy accompanied with neurotrophic keratopathy and tear film instability observed clinically agrees with the large keratinized cells in the superficial corneal epithelium by in vivo confocal microscopy in these patients. Our findings suggest that the loss of corneal nerves contributes to impairment of corneal sensation in patients with type IV or V HSAN. [ABSTRACT FROM AUTHOR]

Purpose: The aim of the study was to quantify corneal densitometry in patients with Fuchs endothelial dystrophy (FED) after endothelial keratoplasty. Materials and methods: We retrospectively reviewed the charts and anterior segment data of patients with FED before and after endothelial keratoplasty. Patients were examined using the Scheimpflug-based Oculus Pentacam corneal densitometry module. Densitometry parameters in different corneal layers and in different annuli were extracted and analyzed. Results: 27 eyes of 27 patients after endothelial keratoplasty (11 DSAEK, 16 DMEK) were included. After endothelial keratoplasty the total corneal light backscatter at total corneal thickness in the central cornea (0–2 mm annulus) was significantly lower than before (DSAEK:p= 0.026, DMEK:p= 0.001). In the entire group the total corneal light backscatter at total corneal thickness and at total diameter before surgery correlated with the postoperative values (Pearson correlation = 0.49,p= 0.01). The strongest correlation was found in the central layer in the DMEK group (Pearson correlation = 0.79,p< 0.001). Conclusions: Corneal densitometry is a useful, objective method for quantification of the outcome of posterior lamellar keratoplasty irrespective of visual acuity. There is a significant correlation between preoperative and postoperative corneal light backscatter values after endothelial keratoplasty, especially in the case of the DMEK procedure. [ABSTRACT FROM PUBLISHER]

Purpose: To investigate the therapeutic efficacy of tumor necrosis factor (TNF)-α stimulated gene/protein 6 (TSG-6) in a rat model of corneal alkali injury. Methods: Corneal alkali injury was produced by placing an NaOH-soaked filter paper disk on the central cornea of the right eye of an anesthetized male Lewis (LEW/Crl) rat. Recombinant human TSG-6, or an equal volume of phosphate-buffered saline (PBS), was administered intravenously (IV), by anterior chamber (AC) injection, or as a topical drop. The affected eyes were photographed daily using a dissecting microscope and documented for clinical time course analysis of corneal opacification. Corneal tissue was excised at pre-determined therapeutic endpoints, with subsequent qRT-PCR or histological analyses. Results: The continuous monitoring of corneal alkali injury progression revealed TSG-6 treatments do not show sufficient effectiveness in vivo regardless of IV injection, AC injection, or topical application. Corneal opacification and neovascularization were not diminished, and gene expression was not impacted by these treatments. However, both IV and AC administration of TSG-6 significantly suppressed pro-inflammatory cytokines compared to PBS-treated eyes. Conclusion: We conclude that the therapeutic potential of TSG-6 is insufficient in a rat corneal alkali injury model. [ABSTRACT FROM AUTHOR]

Purpose/Aim: Despite their importance in accurate mechanical modeling of the cornea, the depth-dependent material properties of the cornea have only been partially elucidated. In this work, we characterized the depth-dependent out-of-plane Young’s modulus of the central and peripheral human cornea with high spatial resolution. Materials and Methods: Central and peripheral corneal buttons from human donors were subjected to unconfined axial compression followed by stress relaxation for 30 min. Sequences of fluorescent micrographs of full-thickness corneal buttons were acquired throughout the experiment to enable tracking of fluorescently labeled stromal keratocyte nuclei and measurements of depth-dependent infinitesimal strains. The nominal (gross) out-of-plane Young’s modulus and drained Poisson’s ratio for each whole specimen was computed from the equilibrium stress and overall tissue deformation. The depth-dependent (local) out-of-plane Young’s modulus was computed from the equilibrium stress and local tissue strain based on an anisotropic model (transverse isotropy). Results: The out-of-plane Young’s modulus of the cornea exhibited a strong dependence on in-plane location (peripheral versus central cornea), but not depth. The depth-dependent out-of-plane Young’s modulus of central and peripheral specimens ranged between 72.4-102.4 kPa and 38.3-58.9 kPa. The nominal out-of-plane Young’s modulus was 87 ± 41.51 kPa and 39.9 ± 15.28 kPa in the central and peripheral cornea, while the drained Poisson’s ratio was 0.05 ± 0.02 and 0.07 ± 0.04. Conclusions: The out-of-plane Young’s modulus of the cornea is mostly independent of depth, but not in-plane location (i.e. central vs. peripheral). These results may help inform more accurate finite element computer models of the cornea. [ABSTRACT FROM AUTHOR]

Purpose: Influence of orthokeratology on corneal biomechanics is equivocal using Ocular Response Analyzer, ORA. Implementing indentation method, corneal tangent modulus was measured and monitored in short-term orthokeratology. Materials and Methods: Sixteen young subjects with refractive errors between -4D to -5D sphere and astigmatism within -1.50D were recruited. One randomly selected eye wore orthokeratology lens (treatment), and the fellow eye wore conventional rigid gas permeable lens (control). Lenses were worn for 30 and 60 minutes and one night separately with a week of washout period in between. The first two visits were randomly scheduled and before the overnight visit. Eyes were kept closed during all the lens wearing periods. Corneal radius, thickness, and biomechanics (using both ORA and an indentation device) were compared between eyes prior to each visit, and then before and after lens wear. Associations between baseline corneal biomechanics and central cornea from overnight visit were investigated. Results: Corneal parameters were similar in each visit before lens wear. Significant corneal flattening was observed in treatment eyes, and flattening increased with wearing time. Control eyes showed no significant corneal curvature changes. Corneal resistance factor (CRF) reduced by 0.42mmHg (± 0.68mmHg) after 30 minutes of orthokeratology treatment. Corneal hysteresis (CH) reduced by 0.42mmHg (+/- 0.63mmHg) in control eyes from overnight wear. Both eyes showed stable tangent modulus, E, throughout the study. A lower CH (r = 0.51, p = 0.046) and a higher E (r = 0.53, p = 0.037) at baseline was significantly associated with greater corneal flattening along the flattest meridian in treatment eyes. Conclusions: Short‐term orthokeratology had no significant effect on corneal tangent modulus. Changes in CH and CRF could be related to their intrinsic measurement variability. Corneal tangent modulus provided another measure of corneal biomechanics. Long-term study is required to investigate predictive role of corneal biomechanics in orthokeratology. [ABSTRACT FROM AUTHOR]

Purpose: To assess long-term morphological and biometric corneal changes produced by overnight orthokeratology and to examine their recovery after cessation of contact lens wear. Methods: Prospective, single-center, longitudinal trial. Fifteen right eyes with low to moderate myopia underwent overnight orthokeratology for 1 year. The central cornea was examined using a confocal microscope and changes determined in visual acuity, refractive error and corneal topography. Cell counts were performed using both the confocal microscope's software and the image analysis software of the USA Health Institute. All measurements were made during orthokeratology treatment and 1 month after discontinuing treatment. Results: No significant changes in endothelial cell density were observed over time but polymegethism increased significantly and baseline values were not recovered ( p < 0.01). Stromal cell density remained unchanged though numbers of activated keratocytes increased during the study and returned to baseline when lens wear ceased. Basal epithelium cell densities significantly fell ( p < 0.01) and epithelial wing and superficial cells showed enhanced visibility ( p < 0.05). Superficial cells increased in height and width; this width increase being significant after 1 year of orthokeratology ( p < 0.01). All epithelial cell changes returned to baseline. Corneal thickness, Bowman layer thickness, subbasal plexus thickness and epithelial thickness were reduced in the central cornea but the stroma was thickened. Of these changes, the decrease in epithelium thickness reached statistical significance ( p < 0.01) and baseline values were not recovered. Conclusion: Overnight orthokeratology induces structural and optical changes particularly in the central corneal epithelium during myopia treatment. If confirmed, the irreversible changes detected indicate a need for further investigation. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the influence of positional deviations on the rebound tonometer measurement. Materials and Methods: Ten white rabbits were used. A bottle of balanced salt solution was connected to the anterior chamber, and the IOP was altered by changing the height of the bottle. A pressure transducer was also connected to the anterior chamber. The IOP was measured by the rebound tonometer (RBT), the Tonopen XL, and the Perkins handheld applanation tonometer at optimal conditions and compared each tonometer. Then, the effect of placing the RBT probe at different distances, different lateral positions, and different angles to the cornea on the IOP was determined. Results: The RBT had the lowest variance in all three tonometers. The IOP measured with the RBT was not significantly altered when the distance from the cornea was 4 to 8 mm, or when the probe was displaced 2 mm to the central cornea, or when the probe was placed at an angle of 20°° from the central axis. Conclusions: The low variance of the RBT measurements and the non-significant changes in the IOP when the probe of the RBT was not on the central axis indicate that the RBT is useful for animal models of glaucoma. Further studies examining its use (repeatability and reliability) in human infants whose eyes and head are usually moving are required. [ABSTRACT FROM AUTHOR]

Purpose: To observe the relationship between the expression of matrix metalloproteinases (MMP-2, MMP-9), heparanase (HPSE), and fibroblast activation protein (FAP) in stroma and corneal neovascularization induced by different etiological factors. Methods: Five models were established: alkaline burn, fungal infection, suturing, immunogen implantation, and tumor cell implantation. The ingrowth time and morphology of corneal neovascularization in each model was observed by slit lamp. Inflammation and neovascularization in the corneal stroma were examined by histopathology. MMP-2, MMP-9, HPSE, and FAP were detected by immunohistochemistry or double immunofluorescence staining. Results: The neovascular vessels started to invade the cornea from the third day in each model. The corneal neovascularization presented dendritic-form, brush-form, and triangle-form in alkaline burn, fungal infection, and suturing models, respectively, and reached to the central cornea in the latter two models. The inflammatory cells appeared in the stroma on the first day, while neovascular vessels grew into the stroma from the third day and both of them accompanied each other from 3-14 days in each model. MMP-2, MMP-9, and HPSE appeared before the neovascularization on the first day and accompanied it from 3-14 days in each model. FAP+ cells occurred mainly around CD31+ vascular endothelial cells in each model. Conclusion: The corneal neovascularization induced by different etiological factors have different morphologies. The inflammation and the expression of MMP, HPSE, and FAP in stroma may serve as pioneers for the growth of corneal neovascularization. [ABSTRACT FROM AUTHOR]

Purpose of the study: We used a rabbit model infected with high phenotypic reactivators (HPRs) as well as recombinant HSV-1 (herpes simplex virus 1) with deletions to study their effect on corneal innervations after latency was established. Materials and methods: Corneas from noninfected New Zealand white rabbits were used to obtain the entire map of corneal innervation. Others were inoculated with the HSV-1 strains McKrae, 17Syn+, or recombinant mutants with glycoprotein K (gK) deletion, or with infected early protein 0 (ICP0) deletion. The animals were euthanized at 124 to 125 days postinfection and the corneas were immunostained with a mouse monoclonal anti-βIII tubulin antibody. Images were acquired with a fluorescence microscope and corneal sub-basal nerve density was calculated on the basis of the whole mount images. Differences between the HSV-infected eyes, and comparison with normal control, were analyzed. Results: In the noninfected rabbit, the stroma was densely innervated in the central area and as a consequence the sub-basal epithelial nerve bundles were shorter, and no vortex was found. The HSV-infected corneas showed nerve damage in both epithelial and stromal nerves. Corneas infected with ICP0 and gK deletion mutants showed mild to moderate damage, while those infected with 17Syn+ and McKrae strains were seriously damaged. In the eyes infected with ICP0 and gK deletion, there were reduced numbers of sub-basal nerve bundles, but most of the corneas retained a normal stromal network. Corneas infected with 17 Syn+ and McKrae displayed destroyed nerve structures and formation of a scar tissue in the central cornea, in which only a few nerve fibers could be detected. Conclusion: HSV-1 primary corneal infection seriously damages the corneal nerves, persisting for more than 4 months. Reduction of axonal transport (by gK deletion) or virus replication (by ICP0 deletion) significantly attenuated the nerve damage induced by the virus. [ABSTRACT FROM PUBLISHER]

Purpose: The corneal subbasal nerve plexus (SNP) offers high potential for early diagnosis of diabetic peripheral neuropathy. Changes in subbasal nerve fibers can be assessedin vivoby confocal laser scanning microscopy (CLSM) and quantified using specific parameters. While current study results agree regarding parameter tendency, there are considerable differences in terms of absolute values. The present study set out to identify factors that might account for this high parameter variability. Materials and methods: In three healthy subjects, we used a novel method of software-based large-scale reconstruction that provided SNP images of the central cornea, decomposed the image areas into all possible image sections corresponding to the size of a single conventional CLSM image (0.16 mm2), and calculated a set of parameters for each image section. In order to carry out a large number of virtual examinations within the reconstructed image areas, an extensive simulation procedure (10,000 runs per image) was implemented. Results: The three analyzed images ranged in size from 3.75 mm2to 4.27 mm2. The spatial configuration of the subbasal nerve fiber networks varied greatly across the cornea and thus caused heavily location-dependent results as well as wide value ranges for the parameters assessed. Distributions of SNP parameter values varied greatly between the three images and showed significant differences between all images for every parameter calculated (p < 0.001 in each case). Conclusions: The relatively small size of the conventionally evaluated SNP area is a contributory factor in high SNP parameter variability. Averaging of parameter values based on multiple CLSM frames does not necessarily result in good approximations of the respective reference values of the whole image area. This illustrates the potential for examiner bias when selecting SNP images in the central corneal area. [ABSTRACT FROM AUTHOR]

Purpose: Ocular injuries following exposure to the chemical agent sulfur mustard (SM) are characterized by acute corneal erosions and inflammation of the anterior segment that may be followed by delayed Partial Limbal Stem Cell Deficiency (LSCD), expressed clinically by corneal neovascularization and epithelial defects. LSCD may derive from direct destruction of limbal stem cells or indirectly from altered limbal stromal niche. The aim of this study was to investigate the mechanism underlying LSCD in SM injuries, focusing on the effects of the chemical on limbal epithelium. Methods: Rabbit eyes were exposed to SM vapor and were observed by slit lamp examinations and pachymetry. Eyes were taken for histological and molecular biology evaluations at different time points (4 h-4 weeks), to include acute and delayed injuries. Epithelial stem cells were identified by ABCG2, p63 and by in vivo BrdU labeling for slow cycling cells. Results: Limbal stem cells were not damaged during the acute phase following SM exposure, in contrast to the severe injury of the central corneal epithelium. On the contrary, limbal epithelium became activated, responding to corneal insult with a wound healing process, as shown by histology and by transient elevation of the stem cells markers. Simultaneously, inflammation was taking place in the limbal stroma lasting for weeks. A gradual loss of stem cells was observed later-on (2-4 weeks), associated with typical symptoms of LSCD. Conclusions: LSCD associated with SM ocular toxicity was not derived from a direct cytotoxic effect on the epithelial stem cells, but apparently from pathological events at the limbal stroma, that produced an abnormal microenvironment for the stem cells, triggering their gradual death. The results, and in particular the absence of a primary damage to the epithelial stem cells, indicate the presence of a therapeutic window for intervention to avoid the development of the delayed LSCD. [ABSTRACT FROM AUTHOR]

Aim and scope: To investigate the inhibitory effect of subconjunctival application of VEGF antibodies bevacizumab, ranibizumab, pegaptanib, and HER2 antibody trastuzumab on corneal neovascularization in a rat model of experimental corneal neovascularization. Material and method: Thirty male Wistar albino rats were included in the study. A chemical burn was induced in central cornea of one eye of the rats by a 75% silver nitrate and 25% potassium nitrate stick. Rats were randomly divided into five groups so that each group contained 6 subjects. Right after the chemical burn, 0.1 ml serum physiologic was injected subconjuctivally in control group (group 1). 1.25 mg/0.05 ml bevacizumab was injected in group 2; 1.2 mg/0.1 ml trastuzumab was injected in group 3; 0.5 mg/0.05 ml ranibizumab was injected in group-4; and 0.3 mg/0.1 ml pegaptanib was injected in group 5. On the 8th day of the experiment, rat corneas were photographed by digital photo-camera. Later, eyes of the sacrificed rats were enucleated and corneal speciements were histopathologically analyzed. The percentages of neovascularization on corneal photographs were examined with digital image analysis. Results: The percentage of corneal neovascularization in all treatment groups was found to be significantly lower than the control group ( p < 0.05). Bevacizumab was found to be more effective than all other agents ( p < 0.05). While the degree of inflammation and vascularization in bevacizumab and trastuzumab groups were significantly lower than the control group ( p < 0.05), the difference was not significant in ranibizumab and pegaptanib groups ( p > 0.05). In all treatment groups, fibroblast intensity was significantly lower than the control group. In terms of corneal thickness, no significant difference was observed between treatment and control groups ( p > 0.05). Conclusion: Bevacizumab, ranibizumab, pegaptanib, and trastuzumab were found effective for the inhibition of corneal NV. In our study we detected that the most effective agent was bevacizumab. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the effectiveness of atelocollagen for canine corneal wound healing. Materials and Methods: Atelocollagen was used to fill a transplant bed in the central cornea, which was then covered with a contact lens. The wound healing process was analyzed clinically, morphologically, and biochemically. Results: At the early healing stage, both the pupillary zone and details of the iris were observed. The stromal collagen fibrils normalized in a time-dependent manner. Type III collagen in the wound area was detected faintly throughout the experimental period. Conclusions: This novel method is advantageous for accelerating wound healing without causing inflammation. [ABSTRACT FROM AUTHOR]

Purpose: To test the hypothesis that anatomical and immunological changes are present in the cornea of patients affected by pterygium. Materials and Methods: The central cornea of 20 eyes of 18 patients with primary pterygium was examined by in vivo confocal microscopy using a 40× lens and an axial resolution of 5 μ m. The size of pterygia was measured by analyzing photographic images. Data from 20 age-matched normal subjects were used as control for analysis. Results: The images obtained showed a significant lower number of epithelial cells in patients with pterygium compared to controls, with superficial epithelial cell area considerably higher than normal, reduced nucleus/cytoplasm ratio, halos around the nuclei, and sharp borders. Numerous highly reflective dendritic-like cells were present in the epithelial cell basal layer. The stroma showed a loss of keratocytes and the presence of lacunae. Tortuosity and number of vesicles of sub-basal nerves were significantly higher than controls. The endothelial cell count showed a normal density of cells of this layer. Conclusions: In vivo confocal microscopy may be helpful in evaluating the immunological and structural changes of the cornea in patients with pterygium and understanding its pathophysiology and the possible role of anti-inflammatory therapy. [ABSTRACT FROM AUTHOR]

Purpose. To investigate inter- and intra-system variations and the effect of corneal drying using a recently developed pDEM. Methods. pDEM was used to simulate "normal" and "dry eye" conditions using two "lacrimation-blink" intervals (20s and 60s). Corneas were examined"graded with sodium fluorescein before and after the experiment. At the end of each experiment. corneas were assessed by trypan blue exclusion technique. Two duplicated pDEM systems were set up and tested to investigate reproducibility. Results. There was no significant difference in the results produced by the two pDEM systems. In the eyes under "normal' condition, there was no significant increase in the fluorescein grading. However, in eyes under "dry eye" condition. fluorescein staining increased and the number of' non-viable cells in the central cornea increased. Conclusions. This novel pDEM system provides a useful assessment tool for the study of causative factors and new treatment strategies for dry eye syndrome. [ABSTRACT FROM AUTHOR]

Purpose. In the limbo-corneal epithelium the stem and early precursor epithelial cell pool is confined to the limbal rim. Among the features associated with this spatial segregation is the general paucity of connexin43 (Cx43) within the limbal basal cell population and its complete absence in resident stem cells. The limbo-corneal epithelial lineage derives from a Cx43-positive (Cx43[sup +]) embryonic outer ectoderm. Accordingly, as a means of identifying the process through which limbal cell phenotypes emerge, we investigated the expression of Cx43 in the ocular surface of embryonic rats. Methods. Ocular surface expression of Cx43 or K12 was determined in cryostat sections of rat embryos and eyes using immunohistological methods. Results. Changes in Cx43 expression revealed the early phenotypic divergence of three main epithelial cell phenotypes of the ocular surface. An analysis of the level and distribution pattern of Cx43 puncta lead to the identification of two distinct domains by embryonic day 10 (El0), a stage that occurs soon after formation of the lens vesicle. Additionally, at E12, ectodermal cells directly adjacent to the edges of the developing retina no longer express connexin. A comparison of anatomical and expression changes throughout embryonic development demonstrated that the two early zones represent the rudiments for the epithelia of the central cornea and conjunctiva, respectively, and that the isolated Cx43-negative (Cx43[sup -]) cells represent the precursors of the basal and, putatively, stem cells of the limbal epithelium. Conclusions. Changes in Cx43 expression revealed that the phenotypic divergence of ocular surface epithelial cells and the generation of limbo-corneal stem cell precursors takes place at a very early stage in ocular development, ahead of the establishment of any identifiable anatomical or differentiation features for these domains. [ABSTRACT FROM AUTHOR]

Purpose. We previously reported that extracellular matrix (ECM) accumulation in human capsular opacification included collagen types I, III, IV, V, and VI. To further characterize the ECM in capsular opacification we performed immunohistochemistry to localize collagen types XII and XIV (fibril-associated collagens with interrupted triple helices, or FACITs) in specimens of human capsular opacification and in cultures of bovine lens epithelial cells (LECs). Methods. Cryosections and paraffin sections of human capsular opacification specimens or uninjured lens capsules, as well as cultured bovine LECs, were processed for immunohistochemistry using antibodies against collagen types I to VI, XII, and XIV. A rat crystalline lens was punctured through the central cornea and the eye was processed for immunohistochemistry for FACITs after healing intervals. Results. In the absence of injury human LECs were unstained for FACITs, but as early as 10 days after operation, LECs in healing capsules were immunoreactive. Collagen types I, III, IV, V, and VI were also detected. ECM deposited in confluent LEC cultures stained for FACITs. Normal rat LECs were not stained for FACITs, but ECM accumulated in injured lens stained for them. Conclusions. LECs up-regulate FACITs during post-opera-tive healing. FACITs, as well as other collagen types, are deposited in ECM in healing injured rat lens, in human capsular opacification and in LEC cultures. ECM components may regulate LEC behavior during postoperative healing. [ABSTRACT FROM AUTHOR]

Purpose. Topical or intracameral administration of H-7 doubles outflow facility and reduces intraocular pressure in cynomolgus monkeys, by relaxing and expanding the trabecular meshwork (TM) and Schlemm's canal (SC). Since H-7 may have anti-glaucoma potential, we determined its effects on the corneal endothelium and ciliary epithelium for safety considerations. Methods. Following topical H-7, aqueous humor flow (AHF), corneal endothelial transfer coefficient (k[sub a]) and anterior chamber (AC) entry of i.v. fluorescein were measured by fluorophotometry; AC aqueous protein concentration ([Protein][sub AC]) was determined by Lowry assay; and corneal thickness and endothelial cell density and morphology were measured by ultrasonic pachymetry and specular microscopy respectively. Following intracameral H-7, specular and/or light and electron microscopy of the corneal endothelium or ciliary epithelium were performed. Results. Following unilateral topical H-7: (1) AHF and k[sub a] were essentially unchanged at 0.5-3.0, 3.5-6.0, and 0.56.0 hr, with an insignificant increase from 0.5-1.5 hr; (2) [Protein][sub AC] was insignificantly increased at 1-1.5 hr but had returned to baseline by 2.5 hr; (3) entry of i.v, fluorescein into aqueous or cornea was modestly and transiently increased; (4) the central cornea thickened significantly at 1-2.5 hr, gradually returning to baseline 2.5 hr after H-7, while peripheral corneal thickness was less affected; (5) corneal endothelial cell borders became indistinct by l hr, but cell morphology was recovering by 3-5 hr and had completely returned to normal by 24 hr; (6) corneal endothelial cell density was unchanged at 5-24 hr. Following intracameral H7, no significant changes were observed in corneal endothelial cell density or morphology by specular microscopy, nor in corneal endothelial or ciliary epithelial morphology by light and electron microscopy. Conclusions. A facility-effective intracameral dose of H-7 had no discernible... [ABSTRACT FROM AUTHOR]

Purpose: The objective is to analyze the antiangiogenic mechanism of suramab, a pharmaceutical compound of bevacizumab and suramin, in a rabbit model of corneal angiogenesis. Material and Methods: Corneal neovascularization was induced in four groups of six New Zealand White rabbits by applying a filter paper disk soaked in 1 M Na (OH) on the central cornea. Group one was treated after injury with intravenous suramab at a dose equivalent to 3 mg/kg of bevacizumab and 10 mg/kg of suramin. Group two was treated with intravenous bevacizumab (5 mg/kg). Group three was treated with 10 mg/kg of suramin while the control group received no treatment. Digital photographs were taken at days 9, 15, 21, and 35. Neovessel formation was quantified giving a 0-4 score to each quadrant according to the centripetal growth of the longest vessel (neovessel index, NVI). Animals were sacrificed at day 35. Corneas were processed for histology, immunohistochemistry, and Western-blot using primary antibodies against P2X2, basic fibroblast growth factor (bFGF), LYVE-1, PECAM-1, and vascular endothelial growth factor-A (VEGF-A). Results: Suramab significantly reduced neovessel growth (mean NVI: 4.2) compared to bevacizumab (8.4), suramin (7.22), and control animals (12.2) at 35 days post-injury (p < 0.01). A lower protein expression of P2X2, bFGF, LYVE-1, PECAM-1, and VEGF-A was found in the cornea of suramab animals than in the other groups of animals. Conclusions: Joint downregulation of bFGF, P2X2, bFGF, and LYVE-1 constitutes a mechanism that induces greater and longer inhibition of corneal angiogenesis. Results might be relevant to ophthalmic care. Ocular administration of suramab is currently being investigated. [ABSTRACT FROM AUTHOR]