Sensory Nerve Retraction and Sympathetic Nerve Innervation Contribute to Immunopathology of Murine Recurrent Herpes Stromal Keratitis.

Purpose: Herpes stromal keratitis (HSK) represents a spectrum of pathologies which is caused by herpes simplex virus type 1 (HSV-1) infection and is considered a leading cause of infectious blindness. HSV-1 infects corneal sensory nerves and establishes latency in the trigeminal ganglion (TG). Recently, retraction of sensory nerves and replacement with "unsensing" sympathetic nerves was identified as a critical contributor of HSK in a mouse model where corneal pathology is caused by primary infection. This resulted in the loss of blink reflex, corneal desiccation, and exacerbation of inflammation leading to corneal opacity. Despite this, it was unclear whether inflammation associated with viral reactivation was sufficient to initiate this cascade of events. Methods: We examined viral reactivation and corneal pathology in a mouse model with recurrent HSK by infecting the cornea with HSV-1 (McKrae) and transferring (intravenous [IV]) human sera to establish primary infection without discernible disease and then exposed the cornea to UV-B light to induce viral reactivation. Results: UV-B light induced viral reactivation from latency in 100% of mice as measured by HSV-1 antigen deposition in the cornea. Further, unlike conventional HSK models, viral reactivation resulted in focal retraction of sensory nerves and corneal opacity. Dependent on CD4+ T cells, inflammation foci were innervated by sympathetic nerves. Conclusions: Collectively, our data reveal that sectoral corneal sensory nerve retraction and replacement of sympathetic nerves were involved in the progressive pathology that is dependent on CD4+ T cells after viral reactivation from HSV-1 latency in the UV-B induced recurrent HSK mouse model.

Late-onset traumatic dislocation of laser in situ keratomileusis corneal flaps: a case series with many clinical lessons.

PURPOSE: To report three cases of late-onset (7-14 years postoperative) traumatic dislocation of laser in situ keratomileusis (LASIK) flaps with epithelial ingrowth that all had delayed surgical intervention. METHODS: A retrospective, interventional case series of three patients who underwent flap repositioning and mechanical debridement of epithelial ingrowth, all after an initial delayed diagnosis/treatment of dislocated LASIK flap. RESULTS: Visual improvement was noted in all three cases following LASIK flap repositioning and debridement of epithelial ingrowth; patient 1 improved from 20/800 corrected-distance-visual-acuity (CDVA) to 20/20 uncorrected-distance-visual-acuity (UDVA) postoperative (14 years post-LASIK, 5-week interval between injury and surgery), patient 2 improved from 20/50 CDVA to 20/20 CDVA (10 years post-LASIK, 4-month interval between injury and surgery) and patient 3 improved from 20/80 CDVA to 20/60 CDVA (7 years post-LASIK, 14-month interval between injury and surgery). Flap dislocation was not suspected or diagnosed in two patients during the first ophthalmic visit. Postoperative visual outcome was not influenced by the presenting vision but might be negatively affected by the delay in surgical intervention, the presence of preoperative central epithelial ingrowth and postoperative striae. CONCLUSIONS: This case series reported one of the longest documented intervals between LASIK and traumatic dislocation of LASIK flap with secondary epithelial ingrowth. Delayed diagnosis and management of flap dislocation after corneal trauma may potentially increase the risk of epithelial ingrowth, recalcitrant flap striae and visual impairment. The presence of late-onset epithelial ingrowth in patients with previous LASIK mandates careful examination for occult flap displacement.

Dellen corneal secundario a carcinoma epidermoide de conjuntiva / Corneal dellen secondary to epidermoid carcinoma of the conjunctiva

El dellen corneal es un área de adelgazamiento corneal localizado como consecuencia de la deshidratación provocada por la falta de humectación relacionada con una zona adyacente de abultamiento conjuntival. Se presenta una paciente femenina de 72 años, blanca, con antecedentes de hipertensión arterial, quien refirió lagrimeo y sensación de cuerpo extraño en el ojo izquierdo, de una semana de evolución. Al examen biomicroscópico se observó tejido fibrovascular en conjuntiva bulbar nasal que infiltraba la córnea, sobreelevado, de superficie lisa, no queratinizada, acompañada de inyección conjuntival y múltiples neovasos. En la córnea periférica y adyacente a esta se observó depresión de base limpia, redondeada, de 6 mm de extensión y profundidad hasta estroma anterior. Teniendo en cuenta las características de la lesión, se planteó como diagnóstico presuntivo dellen corneal secundario a pterigium grado II sintomático. La biopsia informó carcinoma de células escamosas, moderadamente diferenciado, asociado con un extenso infiltrado inflamatorio a predominio de linfocitos(AU)

Corneal dellen is an area of localized corneal thinning as a result of dehydration caused by lack of humectation associated to an adjoining conjunctival bulging. Here is a 72-years old Caucasian woman with a history of hypertension, who presented continuous tearing and feeling of a foreign body in her left eye during a week. The biomicroscopic exam showed fibrovascular tissue in nasal bulb conjunctiva, which infiltrated into the cornea, it was raised of flat surface, unkeratinized, accompanied with conjunctival injection and multiple neovessels. In the peripheral cornea and adjacent to it, there was a clean, rounded depression measuring 6 mm of extension and depth up to the anterior stroma. The presumptive diagnosis was corneal dellen secondary to symptomatic grade II pterygium. The biopsy yielded the presence of squamous cell carcinoma, moderately differentiated, and associated with extensive inflammatory infiltrate with predominance of lymphocytes(AU)

Dellen corneal secundario a carcinoma epidermoide de conjuntiva / Corneal dellen secondary to epidermoid carcinoma of the conjunctiva

El dellen corneal es un área de adelgazamiento corneal localizado como consecuencia de la deshidratación provocada por la falta de humectación relacionada con una zona adyacente de abultamiento conjuntival. Se presenta una paciente femenina de 72 años, blanca, con antecedentes de hipertensión arterial, quien refirió lagrimeo y sensación de cuerpo extraño en el ojo izquierdo, de una semana de evolución. Al examen biomicroscópico se observó tejido fibrovascular en conjuntiva bulbar nasal que infiltraba la córnea, sobreelevado, de superficie lisa, no queratinizada, acompañada de inyección conjuntival y múltiples neovasos. En la córnea periférica y adyacente a esta se observó depresión de base limpia, redondeada, de 6 mm de extensión y profundidad hasta estroma anterior. Teniendo en cuenta las características de la lesión, se planteó como diagnóstico presuntivo dellen corneal secundario a pterigium grado II sintomático. La biopsia informó carcinoma de células escamosas, moderadamente diferenciado, asociado con un extenso infiltrado inflamatorio a predominio de linfocitos(AU)

Corneal dellen is an area of localized corneal thinning as a result of dehydration caused by lack of humectation associated to an adjoining conjunctival bulging. Here is a 72-years old Caucasian woman with a history of hypertension, who presented continuous tearing and feeling of a foreign body in her left eye during a week. The biomicroscopic exam showed fibrovascular tissue in nasal bulb conjunctiva, which infiltrated into the cornea, it was raised of flat surface, unkeratinized, accompanied with conjunctival injection and multiple neovessels. In the peripheral cornea and adjacent to it, there was a clean, rounded depression measuring 6 mm of extension and depth up to the anterior stroma. The presumptive diagnosis was corneal dellen secondary to symptomatic grade II pterygium. The biopsy yielded the presence of squamous cell carcinoma, moderately differentiated, and associated with extensive inflammatory infiltrate with predominance of lymphocytes(AU)

Ocular chemical burns from accidental exposure to topical dermatological medicinal agent.

We report a case of accidental ocular chemical injury by self-medication with a single application of a topical ayurvedic medication containing salicylic acid, phenol, and tincture iodine, which is being used in developing countries for treatment of various dermatological conditions.

Loss of tenascin X gene function impairs injury-induced stromal angiogenesis in mouse corneas.

To determine the contribution by tenascin X (Tnx) gene expression to corneal stromal angiogenesis, the effects were determined of its loss on this response in TNX knockout (KO) mice. In parallel, the effects of such a loss were evaluated on vascular endothelial growth factor (VEGF) and transforming growth factor ß1 (TGFß1) gene and protein expression in fibroblasts and macrophages in cell culture. Histological, immunohistochemical and quantitative RT-PCR changes determined if Tnx gene ablation on angiogenic gene expression, inflammatory cell infiltration and neovascularization induced by central corneal stromal cauterization. The role was determined of Tnx function in controlling VEGF-A or TGFß1 gene expression by comparing their expression levels in ocular fibroblasts and macrophages obtained from wild-type (WT) and body-wide Tnx KO mice. Tnx was up-regulated in cauterized cornea. In Tnx KO, macrophage invasion was attenuated, VEGF-A and its cognate receptor mRNA expression along with neovascularization were lessened in Tnx KOs relative to the changes occurring in their WT counterpart. Loss of Tnx instead up-regulated in vivo mRNA expression of anti-angiogenic VEGF-B but not VEGF-A. On the other hand, TGFß1 mRNA expression declined in Tnx KO cultured ocular fibroblasts. Loss of Tnx gene expression caused VEGF-A expression to decline in macrophages. Tnx gene expression contributes to promoting TGFß1 mRNA expression in ocular fibroblasts and VEGF-A in macrophages, macrophage invasion, up-regulation of VEGF-A expression and neovascularization in an injured corneal stroma. On the other hand, it suppresses anti-angiogenic VEGF-B mRNA expression in vivo.

Assessment of Topical Therapies for Improving the Optical Clarity Following Stromal Wounding in a Novel Ex Vivo Canine Cornea Model.

Purpose: To evaluate the effect of topical suberanilohydroxamic acid (SAHA) and 5-methyl-1-phenyl-2[1H]-pyridone (pirfenidone) on the degree of corneal haze in the stromal wounded ex vivo canine cornea. Methods: Twenty-four corneoscleral rims from normal dogs were uniformly wounded with an excimer laser and placed into culture medium with an air-liquid interface. The control group (n = 8) contained placebo-treated corneas. Treatment group 1 (n = 8) received SAHA topically every 6 hours. Treatment group 2 (n = 8) received pirfenidone topically every 6 hours. Each cornea was fluorescein stained and macrophotographed every 6 hours to assess epithelialization rate. All corneas were also macrophotographed weekly to assess optical clarity (haze). Images were analyzed for differences in pixel intensity between wounded (haze) and unwounded (nonhaze) regions, and haze surface area for each cornea was calculated. Results: The mean epithelialization time was 47.25 hours in the control group, 45.00 hours in the SAHA group, and 43.50 hours in the pirfenidone group, revealing no significant difference (P = 0.368). The median difference in pixel intensity between haze and nonhaze areas was 21.5 in the control group, 8.0 in the SAHA group, and 8.0 in the pirfenidone group, which is significant (P < 0.01). The median haze surface area was 12.96 mm2 in the control group, 5.70 mm2 in the SAHA group, and 5.92 mm2 in the pirfenidone group, which is significant (P < 0.01). Conclusions: Stromal-wounded ex vivo canine corneas exhibited greater optical clarity when treated with SAHA and pirfenidone than when placebo treated at 21 days. There was no significant difference in epithelialization rate between groups. Corneal contour was correlated with geographic haze distribution.

Effect of bone marrow and adipose tissue-derived mesenchymal stem cells on the natural course of corneal scarring after penetrating injury.

In the present study, we investigate and compare the efficacy of bone marrow- and adipose tissue-derived mesenchymal stem cell (MSCs) in corneal wound healing. A penetrating injury was created in the right corneas of Wistar rats (n = 40). Ten microliters of phosphate-buffered solution (PBS) containing 2 × 10(5) green fluorescent protein (GFP) labeled bone-marrow-derived MSCs to group 1 (n = 15), 10 µl of PBS containing 2 × 10(5) GFP-labeled adipose-tissue-derived MSCs to group 2 (n = 15), 10 µl PBS was injected into anterior chamber in group 3 (n = 10, control). Corneal opacity scoring, in vivo confocal microscopy, and histopathological evaluation were done at the end of 8 weeks. Immunofluorescence sections were evaluated to detect transplanted cells. Immune staining was performed to measure the expression levels of keratocan, aldehyde dehydrogenase (ALDH) and CD34. The gene expression levels of tumor necrosis factor (TNF-α), the interleukin 6 receptor (IL-6R), interleukin 12b (IL-12b), and transforming growth factor beta (TGF-ß1) was measured on corneas. The establishment of stem cells in the corneas of the transplanted groups was confirmed by immunofluorescence staining. The expression of keratocan, ALDH, and CD34 increased in the transplanted groups (p < 0.05). The density of keratocytes increased significantly in both transplanted groups according to the in vivo confocal microscopy data (p < 0.05). The expression of TNF-α, IL-6R, and IL-12b decreased significantly in the transplanted groups (p < 0.05). Based on our findings, we consider that allogeneic stem cells facilitate the regeneration of corneal stroma and can be a cell source for stromal repopulation in diseased cornea.

Contact Lens-Related "Keratoschisis"-An Unusual Case of Corneal Stroma Lamellar Trauma.

PURPOSE: To describe the clinical, imaging, and histopathological features of a highly unusual, postcontact lens removal scenario in a 29-year-old woman. Most documented cases recover good vision but differ significantly from our case. METHODS: A 29-year-old woman presented to the Eye Casualty Department as she was unable to remove her right soft contact lens after having inadvertently slept wearing soft contact lenses. During removal, extensive axial corneal epithelial sloughing occurred. Examination immediately after lens removal revealed keratitis and hypopyon, and she was administered intensive topical antibiotics. During her treatment course, a raised crescent of edematous tissue was noted centrally. This persisted at 6 weeks after presentation, so exploration under anesthesia and superficial keratectomy were performed. The preoperative optical coherence tomography (OCT) image showed a thickened inferior cornea with a "tent" of epithelium growing over it. The operative findings identified a flap of the cornea, reflected back on itself and a corresponding stromal depression superior to the flap, correlating well with the OCT findings. The flap was removed and sent for histopathological examination. RESULTS: Histology revealed stromal tissue without the Bowman layer. The corneal stroma was scarred and chronically inflamed. Immunohistochemistry for pan-cytokeratins revealed epithelial cells on both sides of the flap, confirming that regenerative epithelial hyperplasia had occurred over the stromal flap and concurring well with the "tent" of epithelium observed on the OCT. CONCLUSIONS: We have described a highly unusual case of an inferiorly displaced stromal lamellar corneal traumatic flap associated with a soft contact lens removal, which we have termed lens-associated keratoschisis.

Mucocele con expansión orbitaria / Mucocele with orbital expansion

El mucocele es una lesión quística benigna, que se desarrolla en el interior de los senos perinasales por la obstrucción de su drenaje natural en el curso de los procesos inflamatorios, traumas y cirugías. Se presentan tres casos con mucocele frontoetmoidal y expansión intraorbitaria, los cuales fueron atendidos en el Hospital Hermanos Ameijeiras entre los meses de febrero a diciembre del a±o 2013. El propósito del presente estudio es el análisis de los resultados terapéuticos obtenidos y determinar la posible influencia de la vía de abordaje utilizada sobre la efectividad terapéutica y la recidiva tumoral en cada uno de estos pacientes(AU)

Mucocele is a benign cystic lesion that emerges inside the perinasal sinuses due to the obstruction of their natural drainage in inflammatory processes, traumas and surgeries. Here are three cases with frontoethmoidal mucocele and intraorbital expansion, which were treated at "Hermanos Ameijeiras" hospital from February to December, 2013. The objective of this study was to analyze the therapeutic results and to determine the possible influence of the approach path on the therapeutic effectiveness and the tumor recurrence in each of the patients(AU)

Mucocele con expansión orbitaria / Mucocele with orbital expansion

El mucocele es una lesión quística benigna, que se desarrolla en el interior de los senos perinasales por la obstrucción de su drenaje natural en el curso de los procesos inflamatorios, traumas y cirugías. Se presentan tres casos con mucocele frontoetmoidal y expansión intraorbitaria, los cuales fueron atendidos en el Hospital Hermanos Ameijeiras entre los meses de febrero a diciembre del año 2013. El propósito del presente estudio es el análisis de los resultados terapéuticos obtenidos y determinar la posible influencia de la vía de abordaje utilizada sobre la efectividad terapéutica y la recidiva tumoral en cada uno de estos pacientes(AU)

Mucocele is a benign cystic lesion that emerges inside the perinasal sinuses due to the obstruction of their natural drainage in inflammatory processes, traumas and surgeries. Here are three cases with frontoethmoidal mucocele and intraorbital expansion, which were treated at Hermanos Ameijeiras hospital from February to December, 2013. The objective of this study was to analyze the therapeutic results and to determine the possible influence of the approach path on the therapeutic effectiveness and the tumor recurrence in each of the patients(AU)

[Clinical analysis of complications of LASIK using the WaveLight FS200 femtosecond laser].

OBJECTIVE: To explore the incidence of intraoperative and early postoperative complications associated with the WaveLight FS200 femtosecond laser-assisted flap creation in LASIK surgery. METHODS: A consecutive series of 400 patients (786 eyes) who underwent LASIK using the WaveLight FS200 femtosecond laser from March 2011 to December 2012 were included in this retrospective study. Intraoperative and early postoperative complications were described and analyzed. RESULTS: The intraoperative complications included suction loss (two eyes, 0.25%), incomplete flap creation (one eye, 0.13%), severe opaque bubble layers (three eyes, 0.38%), anterior chamber gas bubbles (eight eyes, 1.02%), skip lines (12 eyes, 1.53%), and corneal epithelial defects (eight eyes, 1.02%). The early postoperative complications included canal bleeding (eight eyes, 1.02%), diffuse lamellar keratitis (23 eyes, 2.93%), haze (six eyes, 0.76% ), and flap dislocation (one eye, 0.13% ). CONCLUSIONS: Intraoperative and early postoperative complications of LASIK using the WaveLight FS200 were relatively mild. All patients could obtain satisfactory clinical results with proper treatment.

Corneal Fibroblast Migration Patterns During Intrastromal Wound Healing Correlate With ECM Structure and Alignment.

PURPOSE: To assess keratocyte backscattering, alignment, morphology, and connectivity in vivo following a full-thickness corneal injury using the Heidelberg Retina Tomograph Rostock Cornea Module (HRT-RCM), and to correlate these findings with en bloc three-dimensional (3-D) confocal fluorescence and second harmonic generation (SHG) imaging. METHODS: Rabbit corneas were scanned in vivo both before and 3, 7, 14, and 28 days after transcorneal freeze injury (FI), which damages all corneal cell layers. Corneal tissue was also fixed and labeled for f-actin and nuclei en bloc, and imaged using 3-D confocal fluorescence microscopy and SHG imaging. RESULTS: Using the modified HRT-RCM, full-thickness scans of all cell layers were consistently obtained. Following FI, stromal cells repopulating the damaged tissue assumed an elongated fibroblastic morphology, and a significant increase in cellular light scattering was measured. This stromal haze gradually decreased as wound healing progressed. Parallel, interconnected streams of aligned corneal fibroblasts were observed both in vivo (from HRT-RCM reflection images) and ex vivo (from f-actin and nuclear labeling) during wound healing, particularly in the posterior cornea. Second harmonic generation imaging demonstrated that these cells were aligned parallel to the collagen lamellae. CONCLUSIONS: The modified HRT-RCM allows in vivo measurements of sublayer thickness, assessment of cell morphology, alignment and connectivity, and estimation of stromal backscatter during wound healing. In this study, these in vivo observations led to the novel finding that the pattern of corneal fibroblast alignment is highly correlated with lamellar organization, suggesting contact guidance of intrastromal migration that may facilitate more rapid wound repopulation.

Customised simple limbal epithelial transplantation for recurrent limbal stem cell deficiency.

A 4-year-old boy developed unilateral, total limbal stem cell deficiency (LSCD) following an acid injury. A stable ocular surface was achieved with autologous simple limbal epithelial transplantation (SLET), performed 4 months after the injury. Focal areas of conjunctivalisation were noted in the postoperative period. These were addressed using a novel modification of SLET. A very small piece of limbus from the healthy eye was split into multiple pieces, and applied to the bare corneal stroma with fibrin glue after pannus resection. The surface was covered with amniotic membrane. This resulted in a stable surface, and visual acuity improved to 20/50. The donor remained healthy despite two biopsies being harvested. This case demonstrates that early ocular surface reconstruction may be considered in children with LSCD if amblyopia is a concern. Customised SLET allows surgeons to tackle focal recurrences of LSCD effectively, using minimal limbal tissue from the donor site.

Management of long-standing partially torn and flipped laser in situ keratomileusis flaps.

UNLABELLED: We describe 2 cases of traumatized and torn laser in situ keratomileusis (LASIK) flaps, partially flipped anteriorly or posteriorly, fixed for 8 months or 4 months, and accompanied by epithelial ingrowth. The 2 patients had had uneventful bilateral LASIK 6 years and 1 year before the trauma. In Case 1, the anteriorly flipped flap was removed with transepithelial phototherapeutic keratectomy. Next, mitomycin-C 0.04% was applied for 30 seconds. In Case 2, the portion of the flap that was flipped posteriorly and buried under the remaining intact LASIK flap was restored to its original normal position and epithelial ingrowth was removed mechanically with a microcurette. Irrigation with 20% ethanol was performed to inhibit the recurrence of interfacial epithelial ingrowth. The stretched amniotic membrane overlay over the cornea and sclera was sutured tightly to the episclera as the biologic pressure patch for the inhibition of epithelial re-ingrowth. Good visual acuity was restored in both cases. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Regeneration of corneal epithelium utilizing a collagen vitrigel membrane in rabbit models for corneal stromal wound and limbal stem cell deficiency.

PURPOSE: This study was performed to evaluate the potential of a collagen-based membrane, collagen vitrigel (CV), for reconstructing corneal epithelium in the stromal wound and limbal stem cell deficiency (LSCD) models. METHODS: Three groups of rabbits were used in the stromal wound model: CV affixed using fibrin glue (CV + FG group, n = 9), fibrin glue only (FG group, n = 3) and an untreated control group (n = 3). In the LSCD model, one group received CV containing human limbal epithelial cells (CV + hLEC group, n = 2) and the other was an untreated control (n = 1). Gross observation, including fluorescent staining, pathological examination, immunohistochemistry and electron microscopy, was used to evaluate the effect of CV on the corneal epithelium. RESULTS: In the stromal wound model, fluorescent staining showed that epithelial reconstruction occurred as rapidly in the CV + FG group as it did in the control group. The pathological examination proved that the CV supported a healthy corneal epithelium in the CV + FG group, whereas FG led to hypertrophy and inappropriate differentiation of corneal epithelium in the FG group. In the LSCD model, the corneas in the CV + hLEC group showed sustained tissue transparency with good epithelialization, low inflammatory response and reduced neovascularization. However, the control cornea was translucent and showed high amounts of inflammation and neovascularization. CONCLUSION: We have demonstrated that CV supports corneal epithelial differentiation and prevents epithelial hypertrophy, in addition to serving as a scaffold for hLEC transplantation, without complications.