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Purpose: To determine the efficacy of local (subconjunctival and topical) bevacizumab (Avastin) treatment in patients undergoing vascularized high-risk corneal transplantation., Design: Pilot, prospective, randomized, double-blind, placebo-controlled clinical trial conducted at 5 clinical centers in the United States, India, and Brazil., Participants: Patients aged > 18 years undergoing high-risk penetrating keratoplasty, defined as corneal neovascularization (NV) in 1 or more quadrants ≥2 mm from the limbus or extension of corneal NV to the graft-host junction in a previously failed graft., Methods: Patients were randomized to receive subconjunctival bevacizumab (2.5 mg/0.1 ml) or placebo at the time of surgery, followed by topical bevacizumab (10 mg/ml) or topical placebo, administered 4 times per day for 4 weeks., Main Outcome Measure: The 52-week endothelial immune rejection rate., Results: Ninety-two patients were randomized to receive bevacizumab (n = 48) or control (n = 44). The 52-week endothelial rejection rate was 10% in the bevacizumab group and 19% in the control group (P = 0.20). Post hoc, extended follow-up at the lead study site showed an endothelial rejection rate of 3% in the bevacizumab group and 38% in the control group (P = 0.003). Treatment with bevacizumab was found to have a hazard ratio of 0.15 (95% confidence interval, 0.03-0.65, P = 0.01) in a post hoc Cox regression analysis., Conclusions: In patients undergoing vascularized high-risk corneal transplantation, there was no statistically significant difference in the rate of endothelial rejection at 1 year in the bevacizumab treatment group compared with the control group. This study may have been underpowered to detect a difference between treatment groups, and taken together, our data suggest that, in the current trial design, bevacizumab has a positive but not (yet) significant effect on endothelial rejection., (Copyright © 2022 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Objective: To investigate the corneal graft survival and related risk factors of primary penetrating keratoplasty in congenital corneal opacity infants. Methods: It was a retrospective cohort study. Data were collected from forty-two infants (51 eyes) who were aged ≤12 months and diagnosed with congenital corneal opacity in Beijing Tongren Hospital and Beijing Anzhen Hospital from January 1, 2017 to January 31, 2018. The mean age at surgery was (5.7±2.2) months (3-12 months). The mean follow-up duration was (28.6±2.6) months (24-33 months). All the patients underwent penetrating keratoplasty. The status of the corneal grafts and complications were observed and recorded during the regular follow-up. The survival probabilities were estimated by using the Kaplan-Meier and Log-rank test. The graft survival between different influence factors was analyzed by using the χ 2 test. Results: The Kaplan-Meier survival rates for penetrating keratoplasty were 84.3% (43/51) at 6 months, 78.4% (40/51) at 12 months and 60.8% (31/51) at the last follow-up. The presence of corneal neovascularization was significantly correlated with graft failure ( χ²= 5.264, P =0.022). The graft survival differed between eyes receiving combined surgery and mere penetrating keratoplasty and in eyes with varied surgical indications ( P =0.039, <0.01). Increased intraocular pressure (7 eyes, 13.7%) and persistent epithelial defects (7 eyes, 13.7%) were the most common postoperative complications, followed by complicated cataract (4 eyes, 7.8%) and posterior capsule opacification (2 eyes, 3.9%). Conclusions: The graft survival rate was satisfactory following pediatric keratoplasty although it had a tendency to decrease with the follow-up time. Corneal neovascularization was a major risk factor of graft failure. Surgical indications and procedures also had a certain effect on the graft survival.

Purpose: Corneal neovascularization is the main risk factor for graft rejection after high-risk penetrating keratoplasty (PK). Corneal crosslinking (CXL) has been shown to regress pathological corneal blood and lymphatic vessels and to reduce the risk of graft rejection after high-risk PK experimentally in mice. The aim of this work was to analyze whether CXL is also able to regress corneal neovascularization in patients and is a safe procedure in the context of high-risk PK., Methods: This retrospective case series included 5 patients with progressive corneal neovascularization and the need for high-risk PK because of graft rejection and/or keratitis that received CXL and PK between April 2019 and January 2020. CXL was performed before or in combination with PK and the effect of CXL on corneal neovascularization was assessed morphometrically on slit-lamp images. Patients were followed up to determine the incidence of adverse effects and graft rejection., Results: In 1 case, peripheral corneal CXL was performed first as a single procedure, followed by an additional peripheral CXL procedure combined with PK. In all other cases, peripheral CXL was directly combined with PK. No intraoperative or postoperative complications were observed. Peripheral CXL resulted in a reduction of corneal neovascularization (mean reduction of 70.5% ± 22.7%). Revascularization was not observed. All transplants remained clear and without immune reactions (mean follow-up 16.4 ± 14.9 weeks, range 4-42 weeks)., Conclusions: CXL is able to reduce pathological corneal neovascularization and might therefore be a novel treatment option to improve graft survival after high-risk PK., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: This article explores the application of optical coherence tomography angiography (OCTA) in assessing corneal neovascularization (CoNV) and investigates the features of CoNV in eyes with corneal transplantation., Methods: A pilot, case series, observational study was conducted to enroll patients who underwent corneal transplantation including penetrating keratoplasty (PKP) and deep lamellar keratoplasty (DLKP) with or without additional keratolimbal allograft transplantation. All patients were followed with a series of ophthalmologic examinations including slit-lamp photography and were then imaged with the anterior segment OCTA., Results: The study included 15 eyes of 14 patients (12 men; mean age of 37.4 ± 13.3 years), of which 9 eyes had undergone PKP and 6 eyes DLKP. OCTA was able to clearly identify the features of CoNV in eyes with significant CoNV and to confirm the presence of CoNV in eyes suspected of having CoNV. Four types of CoNV (superficial, stromal, fringe, and recipient-bed CoNV) were discovered by OCTA based on their location and depth. Superficial CoNV was mainly discovered in eyes that had undergone PKP (88.9% ± 11.1%), whereas the recipient-bed CoNV, once thought to be located in the host-graft interface, was discovered to grow in the host cornea only in eyes that had undergone DLKP (83.3% ± 16.7%). Comparing the assessment by 2 masked observers revealed a kappa value of 0.94, indicating excellent agreement., Conclusions: OCTA can be useful to visualize CoNV, which may be valuable in assessing corneal graft rejection.

Purpose: To investigate the anti-(lymph)angiogenic and anti-inflammatory effects of albendazole and to study whether these effects are additive with bevacizumab therapy in a murine corneal suture model. Methods: Corneal neovascularization (NV) and lymphangiogenesis (LY) were compared in a corneal suture model after administration of a subconjunctival injection of albendazole, bevacizumab, dexamethasone, or phosphate-buffered saline (PBS). Immunohistochemical staining and analysis were performed in each group. Real-time polymerase chain reaction (RT-PCR) was performed to quantify the expression of inflammatory cytokines (tumor necrosis factor [TNF]-alpha and interleukin-6), vascular endothelial growth factor (VEGF)-A, VEGF-C, vascular endothelial growth factor receptor (VEGFR)-2, and VEGFR-3. To evaluate the additive effect of albendazole, corneal NV and LY were also analyzed in a combined group of albendazole and bevacizumab therapy and the additive effect was compared with that in the group of double dose of bevacizumab. Results: The albendazole group showed less NV and less LY compared with the PBS control group ( P  < 0.01). When albendazole was combined with bevacizumab therapy, a significant decrease in NV and LY was seen compared with bevacizumab treatment alone, and with albendazole alone (all P values <0.05). The combination group showed better antilymphangiogenesis effect than the group of double dose bevacizumab. The albendazole-treated group showed reduced expression of VEGF-A, VEGF-C, TNF-alpha, and VEGFR-2 compared with corneas from the PBS group ( P value <0.05 in all respective comparisons). Conclusion: Albendazole significantly decreased NV and LY in the cornea. This beneficial effect is additively enhanced when combined with bevacizumab treatment.

Purpose: To investigate the anti(lymph)angiogenic and anti-inflammatory effects of 0.5% timolol maleate in a murine corneal suture model., Methods: Corneal neovascularization and lymphangiogenesis were compared in groups of mice that underwent corneal suture and were subsequently administered a subconjunctival injection of 0.5% timolol maleate, dexamethasone, or phosphate-buffered saline (PBS). Immunohistochemical staining and analysis were performed in each group. Real-time polymerase chain reaction (RT-PCR) was performed to quantify the expression of inflammatory cytokines [TNF-alpha and interleukin (IL)-6], vascular endothelial growth factor (VEGF)-A, VEGF-C, vascular endothelial growth factor receptor (VEGFR)-2, and VEGFR-3., Results: When corneas from the timolol-treated group were compared to the PBS-treated group, we observed decreases in angiogenesis, lymphangiogenesis, and inflammatory infiltration in the timolol-treated group (P value <0.05 in all respective comparisons). Corneas from the timolol-treated group showed reduced expression of VEGF-A, VEGF-C, TNF-alpha, IL-6, VEGFR-2, and VEGFR-3 compared to corneas from the PBS group (P value <0.05 in all respective comparisons)., Conclusion: Blocking adrenergic signaling in the cornea with 0.5% timolol maleate decreased corneal neovascularization and lymphangiogenesis.

Purpose: To describe a method to assess corneal neovascular (CoNV) complexes and identify feeder vessels for selective arterial fine-needle diathermy (FND)., Methods: In patients with CoNV, color photography and corneal indocyanine green angiography (ICGA) and fluorescein angiography are performed. After injection of indocyanine green and sodium fluorescein dye, videography and single-frame images of the region of interest are recorded. Videography is used to measure the time to leakage to assess vessel maturity to guide medical treatment and to discern afferent from efferent vessels. Single-frame images are then selected to locate the number of afferent vessels for surgery, which are selectively cut with a 25-gauge marked needle for the application of FND., Results: Angiography using fluorescein and indocyanine green allows the characterization of CoNV based on assessment of both morphologic (ICGA) and functional (fluorescein angiography) parameters. The time to leakage of fluorescein dye provides important functional information on vessel maturity and helps discern whether medical treatment should be followed before surgical. ICGA allows the identification and delineation of afferent feeder vessels even in the presence of corneal opacities affecting biomicroscopic visibility. Colocalizing the afferent vessel to a visible venous landmark or branch is helpful for placement of the incision and application of FND. Using the described approach, angiographically identified feeder vessels can be selectively treated by FND with minimal thermal energy applied to the corneoscleral limbus., Conclusions: The described method for angiographically guided assessment of CoNV is a useful approach for guiding the medical and surgical treatment of CoNV.

Purpose: To illustrate the role of corneal angiography in the clinical assessment and surgical treatment of patients with complex corneal neovascularization (CoNV)., Methods: A case series of 3 patients with CoNV is presented whose management was guided by indocyanine green (ICG) and fluorescein corneal angiography. In the first case, there was recurrent lipid exudation into an intrastromal cleft from CoNV; in the second, there was progressive exudation from CoNV at the graft-host interface; in the third, CoNV was associated with rejection after deep anterior lamellar keratoplasty., Results: In the first case, angiography helped to identify and treat the feeder vessels and stop further leakage. In the second case, it was possible using angiography to differentiate CoNV arising from iris and limbal vasculature enabling angiographic-guided fine-needle diathermy with cessation of exudation. In the third case, angiography revealed the location of CoNV in the host-graft interface after deep anterior lamellar keratoplasty, rather than within the corneal stroma., Conclusions: Corneal angiography is a useful diagnostic tool to guide medical and surgical management of CoNV by enabling the localization of vessel depth and topography.

The purpose of our review was to familiarize the readers with the new concepts in ocular surface diseases and reconstruction. Limbal stem cell deficiency is characterized by the progressive invasion of conjunctival epithelial cells onto the cornea, superficial vascularisation, destruction of the corneal basement membrane, and chronic inflammatory cell infiltration. Depending on the severity of the disease and the time passed from the primary injury amniotic membrane transplantation, keratolimbal allograft and autograft are the available treatments hoping that, in the nearest future, stem cell transplantation and tissue engineering will become the usual therapeutic choices.

The extension of blood vessels into the normally avascular stroma defines corneal neovascularization. Though this phenomenon, pathophysiological and clinical features are well characterized, therapeutic modalities have been hindered by a lack of safe, efficacious and non-controversial treatments. In this literature review, we focus on available therapeutic options in light of recent evidence provided by animal and clinical studies. First, this review will focus on pharmacological treatments that target angiogenesis. The low cost and market availability of bevacizumab make it the first anti-angiogenic therapy choice, and it has demonstrable efficacy in reducing corneal neovascularization when administered topically or subconjunctivally. However, novel anti-angiogenic molecules targeting the intracellular pathways of angiogenesis (siRNA, antisense oligonucleotides) provide a promising alternative. Laser therapy (direct photocoagulation or photo-dynamic therapy) and fine needle diathermy also find a place in the treatment of stabilized corneal neovascularization alone or in association with anti-angiogenic therapy. Additionally, ocular surface reconstruction using amniotic membrane graft or limbal stem cell transplantation is essential when corneal neovascularization is secondary to primary or acquired limbal deficiency., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

Purpose: To report the outcomes of corneal fine needle diathermy (FND) with adjuvant intrastromal and subconjunctival bevacizumab injection for corneal neovascularization (CN) in children., Methods: Medical records of all children who had undergone FND with adjuvant bevacizumab injection were reviewed retrospectively. Treatment efficacy was evaluated by changes in visual acuity, regression of CN, and clearing of lipid deposits with the aid of slit-lamp color images that were taken before surgical intervention and at last follow-up visit. Postoperative complications were recorded and served to assess the safety of the procedure., Results: Nine eyes of 9 patients were included in the study. The mean age of the patients was 8.4 ± 4.2 years (4-15 years) and the mean follow-up time was 18.7 ± 12.2 months (5-35 months). Three eyes had a history of herpes simplex keratitis (HSK), 3 eyes had complete corneal anesthesia, 2 eyes had CN following suture tract infection after corneal transplant for HSK scar and limbal dermoid excision, and 1 eye had blepharokeratoconjunctivitis. After treatment, 8 eyes had complete CN resolution, and 1 eye with corneal anesthesia following brain tumor resection had partial regression in vessel distribution and size. Lipid deposition clearance lagged behind CN resolution. Mean duration of CN before treatment was 15.3 ± 14.0 months (1-37 months). Mean corrected distance visual acuity before and after surgery was 0.66 ± 0.31 and 0.50 ± 0.37 logMAR, respectively (P = 0.02)., Conclusions: Corneal FND with adjuvant bevacizumab injection is effective at treating sectorial corneal vessels in children.

The cornea in most species is physiologically avascular, and thus this assay allows the measurement of newly formed vessels. The continuous monitoring of neovascular growth in the same animal allows the evaluation of drugs acting as suppressors or stimulators of angiogenesis. Under anesthesia a micropocket is produced in the cornea thickness and the angiogenesis stimulus (tumor tissue, cell suspension, growth factor) is placed into the pocket in order to induce vascular outgrowth from the limbal capillaries. Neovascular development and progression can be modified by the presence of locally released or applied inhibitory factors or by systemic treatments. In this chapter the experimental details of the avascular cornea assay, the technical challenges, and advantages and disadvantages in different species are discussed. Protocols for local drug treatment and tissue sampling for histology and pharmacokinetic profile are reported.

Purpose: The aim of this study was to describe novel, flap-based tattooing techniques for the treatment of disfiguring corneal scars in blind eyes., Methods: Several new modifications of intrastromal corneal tattooing techniques were performed in 6 patients. In corneas with a low risk of perforation, a large limbus-to-limbus lamellar flap was prepared, and the tattooing dyes were spread over the entire stromal bed. After additional puncturing of the dyes into the stroma, the flap was closed and sutured ("large flap technique"). In fragile corneas where convenient preparation of a large flap was not possible, a central small flap was prepared, and the "pupil" was tattooed in analogy ("small flap technique"). Afterward, the corneal periphery corresponding to the "iris" was tattooed, either by puncturing or injecting the dye into peripheral intrastromal tunnels ("tunnel technique")., Results: Two eyes were tattooed using a large flap, and 4 eyes were tattooed using a small flap. Here, the corneal periphery of 3 eyes was tattooed by puncturing, whereas 1 eye was tattooed using the tunnel technique. All tattooing procedures were performed without complications and with good cosmetic results., Conclusions: These novel, flap-based tattooing techniques are alternatives to previously reported procedures and can be adapted to the individual corneal constitution. Further, the tunnel technique is an easy-to-perform method that provides good tattooing results.

Purpose: The aim of this study was to evaluate the effect of clinical and biochemical effects of subconjunctival bevacizumab injection in eyes with corneal neovascularization (CNV) due to varied etiologies., Methods: This prospective interventional case series included 12 eyes of 12 patients with CNV after failed therapeutic penetrating keratoplasty (4 eyes), viral keratitis (4 eyes), adherent leucoma (2 eyes), and pseudophakic bullous keratopathy (2 eyes). Each eye received 3 doses of 1.25 mg (0.05 mL) of bevacizumab at 1-month intervals. Morphological changes in neovascularization were evaluated at each visit by slit-lamp examination and corneal digital photography. Corneal buttons of 4 eyes that underwent optical penetrating keratoplasty after bevacizumab injections were analyzed for vascular endothelial growth factor (VEGF) expression and were compared with untreated vascularized and normal cadaveric donor corneas., Results: Of the 12 patients, 10 were males and 2 were females. Four eyes received injections in the early phase of vascularization (<12 weeks of onset) of which 3 showed regression of vessels. Eight eyes received bevacizumab in the mature phase (>12 weeks) of which 5 showed regression. The follow-up ranged from 1 to 16 months. Five eyes underwent optical penetrating keratoplasty after receiving the last dose of bevacizumab and maintained graft clarity with no episodes of rejection over a mean follow-up of 9.8 months. Four of these 5 corneal buttons analyzed for VEGF expression revealed clinically significant lower levels of expression as compared with the vascular untreated corneas. No local or systemic adverse effects were noted in any patient., Conclusions: Subconjunctival bevacizumab helps to regress CNV due to a decrease in corneal VEGF levels and might prove beneficial for use in clinical conditions leading to CNV.

Objective: Eye drops of aganirsen, an antisense oligonucleotide preventing insulin receptor substrate-1 expression, inhibited corneal neovascularization in a previous dose-finding phase II study. We aimed to confirm these results in a phase III study and investigated a potential clinical benefit on visual acuity (VA), quality of life (QoL), and need for transplantation., Design: Multicenter, double-masked, randomized, placebo-controlled phase III study., Participants: Analysis of 69 patients with keratitis-related progressive corneal neovascularization randomized to aganirsen (34 patients) or placebo (35 patients). Patients applied aganirsen eye drops (86 μg/day/eye) or placebo twice daily for 90 days and were followed up to day 180., Main Outcome Measures: The primary end point was VA. Secondary end points included area of pathologic corneal neovascularization, need for transplantation, risk of graft rejection, and QoL., Results: Although no significant differences in VA scores between groups were observed, aganirsen significantly reduced the relative corneal neovascularization area after 90 days by 26.20% (P = 0.014). This improvement persisted after 180 days (26.67%, P = 0.012). Aganirsen tended to lower the transplantation need in the intent-to-treat (ITT) population at day 180 (P = 0.087). In patients with viral keratitis and central neovascularization, a significant reduction in transplantation need was achieved (P = 0.048). No significant differences between groups were observed in the risk of graft rejection. However, aganirsen tended to decrease this risk in patients with traumatic/viral keratitis (P = 0.162) at day 90. The QoL analyses revealed a significant improvement with aganirsen in composite and near activity subscores (P = 0.039 and 0.026, respectively) at day 90 in the per protocol population. Ocular and treatment-related treatment-emergent adverse events (TEAEs) were reported in a lower percentage with aganirsen compared with placebo. Only 3 serious TEAEs (2 with aganirsen and 1 with placebo) were considered treatment-related., Conclusions: This first phase III study on a topical inhibitor of corneal angiogenesis showed that aganirsen eye drops significantly inhibited corneal neovascularization in patients with keratitis. The need for transplantation was significantly reduced in patients with viral keratitis and central neovascularization. Topical application of aganirsen was safe and well tolerated., (Copyright © 2014 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Nonlinear multiphoton absorption induced by focusing near infrared (NIR) femtosecond (fs) laser pulses into a transparent cornea allows surgery on neovascular structures with minimal collateral damage. In this report, we introduce an fs laser-based microsurgery for selective treatment of rat corneal neovascularizations (in vivo). Contiguous tissue effects are achieved by scanning a focused laser pulse below the corneal surface with a fluence range of 2.2-8.6 J/cm(2). The minimal visible laser lesion (MVL) threshold determined over the corneal neovascular structures was found to be 4.3 J/cm(2). Histological and optical coherence tomography examinations of the anterior segment after laser irradiations show localized degeneration of neovascular structures without any unexpected change in adjacent tissues. Furthermore, an approximately 30 % reduction in corneal neovascularizations was observed after 5 days of fs laser exposure. The femtosecond laser is thus a promising tool for minimally invasive intrastromal surgery with the aid of a significantly smaller and more deterministic photodisruptive energy threshold for the interaction between the fs laser pulse and corneal neovascular structures.

Corneal transplantation is the oldest and one of the most successful transplant procedures with a success rate in many studies in excess of 90%. The high success rate is mainly attributable to the relatively immune-privileged status of the eye and the fact that the cornea is largely avascular. However, the success rate in patients with failed grafts is much lower such that regrafting is frequently the top indication for corneal transplantation in many centers. Neovascularization is the most important risk factor for rejection, as it allows access of the immune system to the donor tissue, compromising immune privilege of the graft/eye. We have developed a process to modify donor corneal tissue to prevent rejection by a single exposure to a gene therapy vector before surgery (EncorStat(®)). The vector used is based on clinically relevant equine infectious anemia virus (EIAV)-derived lentiviral platform and contains genes for two potently angiostatic genes, endostatin and angiostatin. We show that incubation of rabbit, primate, and human corneal tissue with the EIAV vector mediates strong, stable expression in the corneal endothelium. We have optimized this process to maximize transduction and, once this is complete, maximize the removal of free vector before transplant. Rabbit corneas treated with two different antiangiogenic expression vectors (EIAV-EndoAngio and to a lesser extent EIAV-Endo:k5) significantly suppressed neovascularization in a rabbit model of corneal rejection. As a result, corneal opacity, edema, and inflammatory infiltrates were reduced in these corneas. This study demonstrates that angiogenesis is a suitable target to prevent corneal rejection, and provides the first proof-of-concept data for the development of EncorStat, an ex vivo gene therapy treatment to prevent corneal rejection.

Unlabelled: We report a case of complete regression of severe corneal neovascularization after a triple procedure-phacoemulsification, intraocular lens implantation, and Descemet- stripping automated endothelial keratoplasty., Financial Disclosure: Neither 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.)

Background/aims: Corneal neovascularisation (CoNV) can lead to significant ocular comorbidity with reduction in vision and cosmesis. A number of techniques have been described to reduce CoNV, but these can be expensive. Our study aimed to determine the safety, efficacy and long-term outcomes of fine needle diathermy (FND) for CoNV., Methods: A 5-year retrospective study identified all cases of FND. Indications, intraoperative complications, and postoperative visual acuity, after treatment and retreatments, were documented, along with the procedure time. Evidence of regression and number of retreatments were identified., Results: 56 eyes from 52 patients underwent FND for CoNV. The main indications included herpes simplex keratitis (HSK) (53%, n=25) and microbial keratitis/peripheral ulcerative keratitis (13%, n=6). Pretreatment acuity was significantly correlated with extent of CoNV (p=0.044). One complication was noted during the procedure-an intrastromal and subconjunctival haemorrhage (rate 2%). 68.1% of patients demonstrated regression at first follow-up (mean 6.9 weeks), and 89.3% (n=42) showed regression with two or less treatments. Mean post-FND acuity was 0.72 (range -0.2-3.0) vs 0.82 (-0.2-3.0) preprocedure (p=0.08). VA improved in the HSK subgroup (p=0.012). Mean follow-up was 18.9 months (range 1-56 months)., Conclusions: In the largest case series reported, FND appears to be a safe and effective technique in the long term to induce regression of CoNV, with significant improvement in the VA of patients with HSK.

Purpose: To report the results of Descemet's stripping endothelial keratoplasty (DSEK) for failed therapeutic penetrating keratoplasty (PK)., Methods: Twenty-seven eyes of 27 patients undergoing DSEK for failed therapeutic PK were analysed., Results: The mean age of the patients was 36 ± 13.9 years (range: 14-70 years). The median size of the therapeutic graft was 10 mm (inter-quartile range; IQR 9.5-11 mm). Descemet's membrane stripping was performed in all eyes. Graft clarity was achieved in 20/27 eyes. Six eyes had primary graft failure, and one had interface keratitis in the early postoperative period. In all eyes with primary graft failure, there was progressive stromal vascularization, which led to haziness in the graft. Late postoperative complications were rejection in four eyes, infection in two eyes, secondary graft failure in one eye and vortex keratopathy in one eye. Graft size was found to be a significant risk factor for graft rejection in this series. At the last follow-up visit, the best-corrected visual acuity was ≥20/40 in 5/27 eyes (18.5%), 20/60-20/40 in 12/27 eyes (44.4%), 20/100-20/60 in 3/27 eyes (1.5%) and ≤20/200 in 7/27 eyes (25.9%)., Conclusions: Considering the limited success of repeat PK in failed large therapeutic keratoplasty, DSEK is a viable option for visual rehabilitation in these eyes, however; visual acuity may be limited due to sub-epithelial/stromal or interface scarring., (© 2013 The Authors. Acta Ophthalmologica © 2013 Acta Ophthalmologica Scandinavica Foundation.)

Purpose: This study evaluated the clinical outcomes of a combined method of scraping corneal epithelium, coagulating vessels, and subconjunctival bevacizumab in Descemet stripping automated endothelial keratoplasty (DSAEK) for bullous keratopathy with corneal neovascularization (NV).
, Methods: The study included patients with bullous keratopathy undergoing DSAEK. Indications for DSAEK were advanced pseudophakic bullous keratopathy with superficial and deep corneal vascularization and failed corneal grafts. Patients were treated by scraping the corneal epithelium and lightly coagulating the corneal superficial stromal NV and the feeding vessels in the sclera, with a subconjunctival bevacizumab injection at the end of surgery. Subconjunctival and perilimbal bevacizumab dose of 2.5 mg/0.1 mL/affected quadrant was injected at the site of NV in each patient at the end of surgery. One or 2 injections were applied. At each visit, a full eye examination with photographic documentation was performed. Mean follow-up period was 32 (24-36) months.
, Results: Eight eyes of 8 patients with high-risk corneal transplantation and corneal NV were included in this noncomparative interventional case series. The original corneal NV disappeared in all patients immediately after surgery. No patient in the series had recurrent corneal NV or rejection during at least 24 months of follow-up. 
, Conclusions: . The combination of scraping, coagulating, and bevacizumab injection in DSAEK is an effective method to treat corneal NV in corneal transplantation for bullous keratopathy.

Purpose: To evaluate the efficacy and safety of combined feeder vessel coagulation and topical antiangiogenic therapy using bevacizumab in the treatment of mature corneal blood vessels., Methods: Sixteen eyes of 16 patients with mature corneal neovascularization (NV) due to different underlying corneal diseases underwent fine-needle feeder vessel coagulation by diathermy and were treated postoperatively for up to 4 weeks with topical bevacizumab eye drops (containing 5 mg/mL bevacizumab) 5 times a day. Nine patients received an additional subconjunctival bevacizumab injection at the time of cautery., Results: The mean duration of follow-up was 276 ± 147.3 days (range, 29-464 days). Regression of the feeder vessel was observed in 14 eyes. The vascularized area was reduced significantly (P < 0.05). Combined subconjunctival and eye drop antivascular endothelial growth factor treatment was significantly more effective in reducing the vascularized area compared with antivascular endothelial growth factor eye drop therapy alone (P < 0.05). Five patients (5 eyes) needed a second treatment. Thirteen patients (13 eyes) receiving topical bevacizumab treatment combined with feeder vessel coagulation showed stable visual acuity. Two patients had improved visual acuity. One patient had enlarged area of lipid keratopathy despite successful treatment of corneal NV and thus decreased visual acuity. Overall, there was a nonsignificant improvement of best-corrected visual acuity (P > 0.05)., Conclusions: In this pilot study, fine-needle feeder vessel coagulation combined with topical bevacizumab application for treatment of mature corneal NV seemed to be a well-tolerated new treatment option to regress corneal NV. This may not only improve corneal transparency but also "preconditions" such a cornea for future keratoplasty.

Purpose: To report the visual outcomes and graft survival rates of mushroom keratoplasty for the treatment of postinfectious corneal scars., Design: Prospective, noncomparative, interventional case series., Methods: A microkeratome-assisted mushroom-shaped keratoplasty was performed in 31 eyes of 31 patients with a central vascularized full-thickness leukoma, resulting from infectious keratitis of various origin (herpes simplex virus, n = 16; bacteria, n = 10; Acanthamoeba, n = 5), with healthy endothelium. The donor graft consisted of a large anterior stromal lamella (9.0 mm in diameter and ± 250 μm in thickness) and a small posterior button (5 to 6 mm in diameter). Visual acuity, refraction, and endothelial cell density were evaluated before surgery, as well as at 12, 24, and 36 months after surgery, and the postoperative graft survival rate was evaluated., Results: Three years after surgery, in 26 (83.8%) of 31 patients, best spectacle-corrected visual acuity was 20/40 or better with a refractive astigmatism of 4.5 diopters or less. The endothelial cell count at the last follow-up examination averaged 1584 ± 381 cells/mm(2), with an average cell loss of 40.7% from the preoperative value. The survival rate at 3 years was 90.3%, improving to 96.7% when excluding nonimmunologic causes for graft failure., Conclusions: Similarly to penetrating keratoplasty, microkeratome-assisted mushroom keratoplasty restores vision in eyes with postinfectious, full-thickness, central corneal scars. For these vascularized corneas at high risk for immunologic rejection, mushroom keratoplasty combines the visual and refractive advantages of large penetrating keratoplasty grafts with the high survival rate of small penetrating keratoplasty grafts., (Copyright © 2012 Elsevier Inc. All rights reserved.)

PURPOSE. High-risk corneal transplantation refers to grafting performed on inflamed and highly vascularized host beds. It represents a clinical dilemma because the rejection rate can be as high as 90%, irrespective of current treatment modalities. This study was conducted to investigate whether combined blockade of VEGFR-3 (vascular endothelial growth factor receptor-3) and VLA-1 (very late antigen-1) promotes high-risk transplant survival and how it correlates with corneal lymphangiogenesis and hemangiogenesis before and after transplantation. METHODS. High-risk corneal transplantation was performed between normal C57BL/6 (donor) and inflamed BALB/c (recipient) mice. The recipients were randomized to receive intraperitoneal injections of VEGFR-3 and VLA-1-neutralizing antibodies or their controls twice a week for up to 8 weeks after transplantation. Corneal grafts were evaluated by ophthalmic slit-lamp biomicroscopy and analyzed by Kaplan-Meier survival curve. Additionally, whole-mount corneas before and after transplantation were examined by immunofluorescent microscopic assays, and the correlation between lymphatic or blood vessel distribution and transplant outcome was analyzed. RESULTS. The combined blockade markedly promotes 90% survival of high-risk transplants. This strategy specifically modified host beds by selective inhibition of lymphangiogenesis but not hemangiogenesis. A strong correlation was also identified between high-risk transplant rejection and severe lymphatic invasion reaching the donor-graft border. CONCLUSIONS. These novel findings not only provide a new and potentially powerful strategy to promote high-risk transplant survival, they also confirm a critical role of high-degree lymphangiogenesis in mediating high-risk transplant rejection. Results from this study may also shed new light on our understanding and management of other lymphatic- and immune-related diseases in general.

A 62-year-old female visited our clinic with progressively decreased vision in both eyes beginning 12 years prior. Idiopathic corneal opacity in all layers of the cornea was found in both eyes. One year later, we performed penetrating keratoplasty on the undiagnosed right eye. During post-surgical follow-up, corneal edema and stromal opacity recurred, and penetrating keratoplasty was performed two more times. The patient's total serum protein level, which had previously been normal, was elevated prior to the final surgery. She was diagnosed with monoclonal gammopathy of undetermined significance. We made a final diagnosis of monoclonal gammopathy-associated crystalline keratopathy after corneal biopsy. Monoclonal gammopathy-associated crystalline keratopathy is difficult to diagnose and may lead to severe visual loss. A systemic work-up, including serologic tests like serum protein or cholesterol levels, is needed in patients with unexplainable corneal opacity.

Purpose: To evaluate the performance of structurally reinforced, stabilized recombinant human collagen-phosphorylcholine (RHCIII-MPC) hydrogels as corneal substitutes in a rabbit model of severe corneal damage., Methods: One eye each of 12 rabbits received a deep corneal alkali wound. Four corneas were implanted with RHCIII-MPC hydrogels. The other eight control corneas were implanted with either allografts or a simple cross-linked RHCIII hydrogel. In all cases, 6.25 mm diameter, 350 μm thick buttons were implanted by anterior lamellar keratoplasty to replace damaged corneal tissue. Implants were followed for nine months by clinical examination and in vivo confocal microscopy, after which implanted corneas were removed and processed for histopathological and ultrastructural examination., Results: Alkali exposure induced extensive central corneal scarring, ocular surface irregularity, and neovascularization in one case. All implants showed complete epithelial coverage by four weeks postoperative, but with accompanying suture-induced vascularization in 6 out of 12 cases. A stable, stratified epithelium with hemidesmosomal adhesion complexes regenerated over all implants, and subbasal nerve regeneration was observed in allograft and RHCIII-MPC implants. Initially acellular biosynthetic implants were populated with host-derived keratocytes as stromal haze subsided and stromal collagen was remodeled. Notably, RHCIII-MPC implants exhibited resistance to vascular ingrowth while supporting endogenous cell and nerve repopulation., Conclusions: Biosynthetic implants based on RHC promoted cell and nerve repopulation in alkali burned rabbit eyes. In RHCIII-MPC implants, evidence of an enhanced resistance to neovascularization was additionally noted.

Introduction: Corneal vascularization develops as a result of various corneal diseases. Fine- needle diathermy (FND) is one of the methods to occlude corneal vessels., Objective: To evaluate the efficacy and safety of this method in a series of patients with corneal vascularization., Materials and Methods: Twenty eight patients with corneal vascularization were treated with FND. The patients were divided into four groups according to etiological factors like pre-keratoplasty (high risk patients), increased keratopathy, recurrent inflammation and postkeratoplasty rejection .The FND was performed under local anaesthesia with the help of a corneal needle of 10-0 monofilament nylon suture. The success of treatment was evaluated in terms of regression and occlusion of corneal vessels., Results: The main aim of a complete occlusion of blood vessels was achieved in all patients. The most frequent complication seen was a transient whitening of the cornea especially around the needle entry in nine cases, which cleared within 24 hours., Conclusion: FND is a simple safe, inexpensive and effective treatment. It can be repeated and used as an alternative therapy for corneal vascularization., (© NEPjOPH.)

Purpose: To investigate whether corneal graft survival could be improved by topical or subconjunctival bevacizumab in a murine model of vascularized high-risk corneal transplantation., Methods: Before corneal transplantation, intrastromal sutures were placed for 2 weeks in the corneas of BALB/c mice, inducing intense angiogenesis. Allogeneic corneal transplantation was performed using C57BL/6 donor mice. Topical bevacizumab (2.5%) was delivered 3 times a day for 3 weeks in one treatment group, and 0.02 mL (0.5 mg) bevacizumab was injected subconjunctivally at days 0, 4, 8, and 15 after transplantation in the other treatment group. The control group received no treatment. Grafts were examined twice a week for 8 weeks by slit-lamp microscopy and were photographed once a week by slit-lamp digital camera and scored for opacity. For assessment of corneal neovascularization (NV), a quantitative method was used to measure three primary metrics including neovascular area, vessel caliber, and neovessel invasion area., Results: Both topical and subconjunctival bevacizumab treatment reduced neovascular area and vessel caliber; however, the regression of corneal NV was more profound when treated subconjunctivally. The mean percentage reduction of neovascular area was 55% (P < 0.05) by week 8 in the subconjunctival treatment group and 33% (P = 0.15) in the topical group. Only subconjunctival bevacizumab treatment resulted in significant regression of neovessel invasion area (P < 0.05). All corneal transplants in both the control and the topical groups were rejected by 4 weeks after transplantation. However, in the subconjunctival treatment group, 33% of corneal grafts survived (P < 0.01)., Conclusions: Subconjunctival bevacizumab may offer an adjunctive measure to conventional therapies in preventing graft rejection in high-risk corneal transplantation.

Purpose: Chemokine receptor 7 (CCR7) is a key homing molecule for immune cell trafficking, including corneal antigen-presenting cell (APC) migration from the inflamed cornea to draining lymph nodes (LNs). Here, the authors investigated the effect of CCR7-facilitated donor APC trafficking on allosensitization, regulatory T-cell (Treg) function, and graft survival in corneal transplantation., Methods: CCR7(-/-) or wild-type (WT) allogeneic corneal grafts were transplanted onto the neovascularized high-risk recipient beds. Two weeks later, the frequency of directly alloprimed host T cells was measured by the IFN-gamma ELISPOT assay. Treg function was tested by a coculture suppression assay and an IFN-gamma ELISPOT assay. Kaplan-Meier analysis was performed to evaluate graft survival., Results: The recipients of CCR7(-/-) grafts had fewer migrated donor APCs and lower frequency of IFN-gamma-producing T cells in the draining LNs. However, there was no statistically significant difference in transplant survival between recipients of CCR7(-/-) and those of WT grafts. Tregs from the CCR7(-/-) graft recipient group showed reduced regulatory potential for the suppression of proliferation of naive T cells and direct alloprimed T cells and expressed lower Foxp3 levels. In vitro studies confirmed that mature CCR7(+) major histocompatibility complex class II(+) CD86(+) graft-derived dendritic cells were critical for Treg function., Conclusions: Not only is CCR7-mediated donor-derived APC trafficking to the draining LNs important in the initiation of host T-cell priming, it is crucial for Treg-mediated tolerance.

Purpose: To evaluate the short- and long-term in vivo safety and efficacy of topical bevacizumab (Avastin) application for treatment of corneal neovascularization secondary to a variety of corneal diseases., Methods: Thirty eyes of 27 patients with progressive corneal neovascularisation (not responding to conventional anti-inflammatory treatment) due to different underlying corneal diseases received topical bevacizumab (Avastin) eye drops (5 mg/ml Bevacizumab) for 0.5-12 months (five times/day on average). At each visit, a routine Snellen visual acuity assessment was performed, followed by ophthalmic examination including fluorescein staining. Changes of corneal neovascularization and vessel diameter were assessed using morphometry of standardized digital corneal photographs., Results: Five patients (five eyes) developed new corneal epithelial defects during topical bevacizumab treatment. In 22 patients, no new epithelial defects were observed. None of the 27 patients complained about any drug-related ocular or systemic adverse events during follow-up. No allergic reactions were observed. Corneal photographs of 21 eyes (19 patients) could be assessed. The mean reduction in vascularized area during treatment was 61%. The mean reduction in vessel diameter under topical Avastin therapy was 24%., Conclusions: Off-label topical bevacizumab therapy against corneal neovascularization secondary to different corneal diseases was generally well-tolerated for up to 12 months. Bevacizumab (Avastin) eye drops inhibit corneal neovascularization, and lead to a reduction of the vessel diameter. Our results suggest that off-label use of Bevacizumab eye drops is a relatively safe and well-tolerated option for the treatment of corneal neovascularization. Care should be taken in patients with epithelial defects and neurotrophic keratopathy.

A 68-year-old lady presented with irritation in left eye of one-week duration. Examination of her left eye revealed loose corneoscleral sutures with corneal vascularization and white deposit in the cornea in front of the blood vessels. She underwent laser photocoagulation but later developed necrotizing scleritis. We report a rare complication of necrotizing scleritis following laser photocoagulation of the corneal new vessels.

Purpose: To report the effect of superficial keratectomy combined with subconjunctival bevacizumab injection in 2 cases of corneal neovascularization (NV)., Methods: An interventional case series was undertaken on 2 patients with corneal NV: 1 due to sclerokeratitis secondary to rheumatoid arthritis and the other due to Terrien marginal degeneration. Both patients underwent superficial keratectomy combined with subconjunctival bevacizumab injection (2.5 mg/0.1 mL)., Results: Corneal NV regressed with the surgical removal and showed no signs of recurrence after 3 months of follow-up. Both patients reported dramatic subjective improvement in their vision within 1-2 weeks. Best corrected visual acuity improved in 1 patient., Conclusion: The combination of superficial keratectomy with subconjunctival injection of bevacizumab may offer a new strategy for the treatment of superficial corneal NV.

Purpose: To evaluate the efficacy of autologous corneal epithelial sheet implantation in restoring transparency of rabbit corneas severely injured by alkaline and the effect of photocoagulation in arresting corneal neovessel ingrowth., Setting: Ophthalmology Department, School of Biomedical Sciences, Universidad Austral, Buenos Aires, Argentina., Methods: Limbal stem-cell deficiency (LSCD) was induced in 14 rabbits by alkali burns. A limbal cell biopsy was done in the contralateral eye, and the cells were cultured on a fibroblast feeder layer grown on autologous clotted platelet-poor plasma or commercial fibrin for 21 days. Anterior keratectomy was followed by suturing corneal cell sheets over the stroma. If regrowth of vessels occurred, argon laser photocoagulation was applied to them. Rabbits were killed at 30, 60, 90, 180, and 360 days and the corneas processed for histopathology and inmunohistochemistry., Results: A small (2.5 mm(2)) limbal biopsy achieved stem-cell replication in vitro. Corneal clarity and epithelial defects evolved with a trend toward improvement. There was a significant reduction in corneal neovascularization. Histology showed a multilayered stratified epithelium including several epithelial-like cells with clear cytoplasm in the deepest part. There were no signs of intraepithelial mucin cells on the implanted corneas. Immunohistochemical results showed expression of cytokeratins 3 and 12 in the central corneal epithelium and an absence of cytokeratin 19., Conclusions: Autologous limbal epithelial cell transplantation improved the corneal surface in eyes with LSCD. Photocoagulation of neovessel ingrowth was effective over the 1-year follow-up. Results may facilitate the application of this technique in patients.

Purpose: To investigate the efficacy of photodynamic therapy with verteporfin for the treatment of patients with corneal neovascularization., Design: Prospective interventional case series., Methods: Eighteen eyes of 18 patients with stable corneal neovascularization who were refractory to conventional treatment were treated with photodynamic therapy with verteporfin (6 mg/m(2)). Five patients were treated following penetrating keratoplasty (PK), and two patients were treated before PK. Anterior segment photography was performed before and after treatment. Best-corrected visual acuity (BCVA) and area of corneal neovascularization were measured., Results: At the one-year follow-up, 14 eyes (77.8%) showed a decrease in corneal neovascularization, and nine eyes (50.0%) showed complete vascular occlusion. In five patients who had corneal allograft, complete or partial occlusion was achieved in all eyes. Two patients who underwent subsequent keratoplasty did not manifest allograft rejection or revascularization. Seventeen eyes (94.4%) had stable or improved vision. The mean area of corneal neovascularization significantly decreased from 25.5 +/- 14.2 mm(2) to 14.9 +/- 14.6 mm(2) (P < .01), respectively. No significant complications associated with photodynamic therapy were observed except mild stromal haze in one eye., Conclusion: Photodynamic therapy with verteporfin may be effective for the treatment of corneal neovascularization.

Purpose: To describe a technique of corneal vessel occlusion by using electrolysis-needle cautery., Methods: A prospective case series of three patients., Results: Corneal vessels were successfully occluded in all patients. Vessels remained occluded during the first 8 months post-cautery follow up. Two patients needed repeat cautery at 9 and 10 months respectively. Patients found the procedure comfortable. There was no post-operative induced astigmatism., Conclusion: The technique of ENC is simple, effective and controlled. This technique compares favorably and may prove to be more versatile than Fine Needle Diathermy in the occlusion of corneal vessels that lead to lipid keratopathy.

Background: Corneal ulcers with fornix shortening associated with late stages of cicatrizing trachoma contribute significantly to blindness in many developing countries. We report on the outcome of ocular surface and fornix reconstruction using amnion membrane transplantation., Patients and Methods: From 2001 to 2005, cryopreserved human amnion membrane without mitomycin C was grafted to 25 eyes of 17 patients with trophic corneal ulcers and symblepharon (cicatrizing trachoma: 19 eyes of 14 patients, Stevens-Johnson syndrome: 4 eyes of 2 patients, alkali burns: 2 eyes of 1 patient) in a controlled case series. Follow-up was done up to 6 months., Statistics: Fischer's exact probability test., Results: Of 25 eyes, 9 of 19 eyes with trachoma, 3 of 4 eyes with Stevens-Johnson syndrome, and 2 of 2 eyes with chemical burns showed complete reepithelialization and stromal recovery after 28-35 days (mean: 31+/-2.3 days). The primary success rate of trachoma eyes was not significantly different from the other indications (p=0.256). At 6 months post-op, 15 of 19 trachoma eyes (79%) compared to 2 of 6 non-trachoma eyes (33.3%) had developed a recurrence of symblephara (p=0.0592), and 13 of 15 eyes (86.6%) with a cicatricial trachoma compared to 1 of 6 with non-trachoma diagnosis experienced a recurrence of corneal vascularization (difference nonsignificant: p=0.1752). Persistent long-term reepithelialization was observed only in 1 of 19 trachoma eyes (5.3%) versus 4 of 6 non-trachoma eyes (66.7%, p=0.005); 3 of 19 trachoma eyes with a recurrence of ulcers had perforated after 6 months., Conclusions: Human amnion membrane without mitomycin C can be used for ocular surface reconstruction in selected patients with cicatrizing trachoma. Its efficacy in the long-term rehabilitation of cicatrizing trachoma seems to be limited due to the progressive scarring.

Objective: To investigate whether peripheral corneal neovascularization in bullous keratopathy (BK) is due to conjunctivalization, a sign of limbal stem cell deficiency., Design: Observational case-control study., Participants: Sixteen BK patients., Methods: Patients were divided into 2 groups: BK without peripheral neovascularization [NV(-) group; 5 patients, 5 eyes] and BK with neovascularization [NV(+) group; 11 patients, 13 eyes]. Evidence of conjunctivalization was evaluated by periodic acid-Schiff staining of impression cytology samples from the peripheral vascularized cornea. The 2 groups' durations of disease also were compared. Penetrating keratoplasty (PK) was performed in all 16 cases, and the 2 groups' durations of reepithelialization after PK were compared., Main Outcome Measures: Presence of goblet cells using impression cytology, duration of BK, and duration of postoperative reepithelialization., Results: Goblet cells were found on the peripheral corneal surface in all eyes in the NV(+) group. However, all eyes in the NV(-) group were negative for goblet cells (P<0.0001). Duration of disease was 14.4+/-5.4 months in the NV(-) group and 66.2+/-65.5 months in the NV(+) group (P = 0.030). Duration of postoperative epithelialization was 6.2+/-2.2 days in the NV(-) group and 28.8+/-36.5 days in the NV(+) group (P = 0.046)., Conclusion: Conjunctivalization of the peripheral cornea and delayed postoperative epithelialization in BK patients with NV suggest the presence of limbal stem cell deficiency in such patients. Patients with long-standing disease were found to be more prone to neovascularization. For this reason, early surgery may lead to a better surgical outcome.

Purpose: To determine the differential gene expression of chemokine species and leukocyte infiltration of grafts in the early pre-rejection postoperative period after high-risk (HR) compared to normal-risk (NR) corneal transplantation., Methods: Fully mismatched and syngeneic corneal grafts were performed in NR (avascular) and HR (vascularized) recipient beds of BALB/c murine hosts. Gene expression levels of a panel of chemokines were determined by a multiprobe ribonuclease protection assay system. The profiles of infiltrating cells into the corneal grafts at the same times were determined immunohistochemically., Results: Compared to NR transplantation, HR eyes exhibited significantly higher mRNA levels for macrophage inflammatory protein (MIP)-2 and monocyte chemotactic protein-1 (MCP-1) on day 1, and for eotaxin on days 1 and 3 after transplantation. By day 6 after transplantation, still well before graft rejection, significantly higher levels of RANTES, eotaxin, MIP-1alpha, MIP-1beta, MIP-2, and MCP-1 were detected in HR eyes. The overexpression of MIPs in HR eyes correlated with a significant increase in the number of infiltrating macrophages (p<0.01), and neutrophils (p<0.05) in HR compared to NR recipients. Low levels of eosinophil and mast cell infiltration were observed in all grafts, with a modest increase in mast cell infiltration (p<0.05) in HR compared to NR grafts., Conclusions: These results suggest that increased expression of gene products for select chemokines, in particular those that mediate recruitment of innate immune cells, in the early period after HR corneal transplantation is related to the enhanced leukocytic infiltration of grafts observed in HR keratoplasty.

Purpose: To describe a case with bullous keratopathy and anterior segment inflammation associated with heavy liquids., Design: Observational case report., Methods: Review of clinical and histopathologic changes., Results: A 65-year-old patient underwent a pars plana vitrectomy for a rhegmatogenous retinal detachment. Perfluorodecalin was used as a temporary retinal tamponade. After surgery, bubbles of heavy liquid were noted in the anterior chamber. Fifteen months later, severe corneal edema developed, associated with corneal vascularization and keratic precipitates. Removal of heavy liquid through a paracentesis was attempted but the cornea remained edematous, and a penetrating keratoplasty was performed. In the histopathologic examination inflammatory changes from retention of perfluorodecalin were observed. There was a decompensated cornea with florid bullous keratopathy, inflammatory infiltration with vascularization, and deposition of perfluorodecalin within keratocytes and perivascular macrophages., Conclusion: Presence of heavy liquids in the anterior chamber may be associated with an intense inflammatory response and corneal decompensation.

Objective: This experimental study assessed the usefulness of a rat model of corneal angiogenesis for assessing the angiogenic activity of cerebrovascular malformations (CVMs) over time. It is the first investigation to have focused on dynamic quantification of angiogenesis related to CVMs (i.e., assessing changes over time as opposed to one point in time) and the first to have compared different CVMs and other tissues in this way., Methods: The CVM specimens were from 15 randomly selected arteriovenous malformation (AVM) cases, 15 randomly selected cavernous malformation cases, and 2 venous angioma cases. All the samples came from surgical resections performed in the Marmara University Department of Neurosurgery and the Marmara University Institute of Neurological Sciences between January 1998 and January 2003. Glioblastoma multiforme tissues, normal adult brain tissues, and normal brain artery tissues were used as controls. Tissue from each specimen was implanted in one corneal micropocket in each cornea of a single rat, and the level of angiogenic activity in the cornea was graded (low, moderate, or high) at Days 1 through 9 after tissue implantation. Another set of rats was subjected to the same corneal implantation procedure, and two widely accepted indicators of neovascularization, microvessel counts and vascular endothelial growth factor expression, were assessed at Days 3, 5, 7, and 9 after surgery. The tissue-group results for grade of angiogenic activity, microvessel count, and grade of vascular endothelial growth factor expression were compared., Results: Of the three CVM types, the AVMs showed the highest angiogenic activity, cavernous malformations exhibited some degree of angiogenic activity (less than AVMs but more than normal brain artery tissue), and angiogenesis induction by venous angiomas was comparable to that of normal brain artery tissue. Corneas implanted with the AVM samples showed Grade 1 angiogenesis on Day 2, Grade 2 angiogenesis on Day 4, and Grade 2.5 angiogenesis on Day 8. Cavernous malformation samples caused Grade 1 angiogenesis after Day 5, and grading remained less than 1.5. Venous angioma samples resulted in angiogenesis graded less than 1., Conclusion: The results showed that this rat model of corneal angiogenesis is of value for assessing the angiogenic potential of CVMs and for evaluating details of the neovascularization process over time. Further investigation of CVMs with this model might yield information about how angiogenesis can be modulated, and this could lead to development of biological treatments that inhibit this process.

We studied IL-1alpha level in corneal scars with/without neo-vascularization. A total of 27 patients underwent grafting for corneal scar. Recipients were grouped according to number of vascularized quadrants (0 to IV/IV): none (n = 12), one (n = 5), two (n = 4) and four (n = 6). Recipient corneas were collected during surgery and IL-1alpha measured by immunoassay. Controls were donor corneas unsuitable for transplantation. Graft rejection rate was calculated for each group. Mean IL-1alpha concentration in corneal scars was 6 +/- 3.93 pg/mm3; significantly higher as compared to controls (1.25 +/- 2.03 pg/mm3). IL-1alpha correlated well with amount of blood vessels, except in IV/IV scars: 5.17 +/- 3.65 pg/mm3 for 0/IV; 8.02 +/- 2.51 pg/mm3 for I/IV; 8.27 +/- 3.62 pg/mm3 for II/IV and 4.47 +/- 5.03 pg/mm3 for IV/IV corneal scars. Vascularization of corneal scar is associated with increased IL-1alpha level (in all but highly vascularized scars), indicating that IL-1alpha promotes early stages of vascularization. Graft rejection rate increases in patients with higher vascularization, independently of IL-1alpha level.

Purpose: To determine the epithelial lineage of origin in corneal pannus tissue surgically removed from patients with total limbal stem cell (SC) deficiency., Methods: The lineage of origin of the entire conjunctivalized pannus removed from eight corneas with a diagnosis of total limbal SC deficiency was characterized by anti-keratin (K)-3 and anti-K19 monoclonal antibodies. The protein and mRNA of epithelial outgrowth from segments of five such pannus specimens were analyzed by Western blot and reverse transcription-polymerase chain reaction, respectively., Results: Cross sections of all eight specimens showed a stratified epithelium with goblet cells expressing mucin (MUC)-5AC, and a stroma showing blood vessels and inflammatory cell infiltrates. Immunostaining showed full-thickness expression of K19 in the entire pannus of all eight specimens. Expression of K3 was negative in seven patients, but was sporadically positive in a patient with Stevens-Johnson syndrome. In culture, all five pannus specimens generated a compact, small epithelial cell outgrowth, and except for one, reached confluence in 2 to 3 weeks. The K3/K12 pair was expressed by extracts of cell outgrowth from the control limbal epithelial explant, but not in all five pannus specimens. A 60-kDa band of DeltaNp63 was expressed in the control specimen and in all five pannus specimens. Cell outgrowth expressed K3 transcript in three, but none showed K12 transcript., Conclusions: The resultant epithelial phenotype of the pannus tissue was not corneal, as evidenced by the negative staining to cornea-specific K12 mRNA and protein, but was conjunctival, as evidenced by the presence of goblet cells, the weak expression of K3, and the strong expression of K19. The abundant expression of DeltaNp63 in such conjunctiva-derived epithelium in eyes with total limbal SC deficiency raises doubts as to its validity as a limbal SC marker., (Copyright Association for Research in Vision and Ophthalmology)

Background/aims: To evaluate treatment with high peak power pulse energy by femtosecond ultrashort pulse laser (titanium sapphire laser) delivered at an 800 nm wavelength for corneal neovascularisation using photodynamic therapy (PDT) mediated by indocyanine green (ICG)., Methods: Using a gelatin solid as an in vitro corneal model, the safety of laser power was studied to determine if it degenerated gelatin with or without ICG. The authors then induced corneal neovascularisation in rabbit eyes by an intracorneal suturing technique. Fluorescein angiography was used to evaluate occlusion before PDT and 0, 1, 3, and 10 days after PDT. The authors performed light microscopy with haematoxylin eosin staining and transmission electron microscopy to determine thrombosis formation in the neovascular regions., Results: The threshold of peak laser power density ranged from 39 to 53 W/cm(2). Laser irradiation was started 30 seconds after a 10 mg/kg ICG injection, and all irradiated segments were occluded at 0, 1, 3, and 10 days at 3.8 J/cm(2). Light and electron microscopy documented thrombosis formation in the neovascular region., Conclusion: Femtosecond pulse laser enhanced by ICG can be used for PDT. Because of effective closure of corneal neovascularisation at a low energy level, the high peak power pulse energy of the femtosecond pulse laser might be more efficacious than continuous wave laser for use with PDT.

Background: Toxic epidermal necrolysis (TEN) can lead to ocular surface scarring associated with pain, dry eye symptoms, and decreased visual acuity that often are difficult to treat., History and Signs: A 34-year-old woman was referred to our department two-years after TEN induced by lamotrigin (lamictal(R)). She was complaining of severe visual acuity loss, pain, and dry eye symptoms. Visual acuity was reduced to light perception in the right eye (RE) and to 0.2 in left eye (LE). Basal Schirmer test was 2 mm in RE and 3 mm in LE. With or without therapeutic contact lenses, the patient was experiencing severe discomfort requiring tear supplementation up to 50 - 80 times/day in both eyes., Therapy and Outcome: A free autologous mucosal graft (3.5 x 2.0 cm) was transplanted from the lower lip into the upper RE fornix. Six months after surgery, with therapeutic contact lenses, the need for tear supplementation was markedly reduced to 3 - 4 times/day in RE while it remained unchanged in LE. In RE, slit-lamp examination revealed decreases in the corneal stromal edema and in the diameter of neo-vessels associated with an improvement of visual acuity (counting fingers at 30 cm). Basal Schirmer test values were unchanged. It has to be mentioned that the improvement observed after surgery was markedly dependent on wearing a therapeutic contact lens., Conclusions: Free autologous buccal graft transplantation (with its presumably accessory salivary glands) in association with the use of a therapeutic contact lens can be an efficient approach to treat ocular complication following drug-induced TEN (Lyell's syndrome).