Epi-off-lenticule-on corneal collagen cross-linking in thin keratoconic corneas.

PURPOSE: To evaluate the safety and efficacy of corneal collagen cross-linking (CXL) performed on overlaying a corneal lenticule to thin recipient corneas of progressive keratoconus (KC) patients. METHODS: In this study were enrolled eyes of patients affected by progressive KC with a minimum corneal thickness less than 400 µm, after overlaying a lenticule of human corneal stroma prepared with the femtosecond laser. The lenticules used were 100 µm thick and of 8.5 mm diameter in all the cases. Both the host cornea and the lenticules were subjected to epithelial debridement. CXL was carried out according to the standard protocol. Visual acuity, refraction, slit-lamp examination, endothelial cell density, pachymetry and keratometry, anterior segment tomography (AS-OCT) and confocal microscopy were evaluated preoperatively and at 1, 3, 6 and 12 months postoperatively. RESULTS: CXL was performed in 10 eyes of 8 patients (main age 23), corneal thickness range 379-414 µm, mean 387.6 µm. One patient was lost at follow-up. In all other cases, visual acuity and the endothelial cell density remained stable over a 12-month follow-up. Preoperative mean K1 and mean K2 were 46.91 ± 1.9 and 50.75 ± 2.93, respectively, and at 12 months mean K1 was 47.36 ± 2.66 and mean K2 50.53 ± 3.35. The AS-OCT clearly showed a demarcation line in all patients at 1, 3 (mean depth 283 µm and 267 µm, respectively) and in some cases at 6 months. Reduced keratocyte density and stromal oedema were observed immediately up to 1 month after treatment, while a slight subepithelial haze was present at 1-month and completely disappeared by 6 months. CONCLUSION: This new technique seems to offer a therapeutic opportunity for young patients suffering from progressive KC with very thin corneas, in which the standard treatment is not indicate, and delay or avoid the need for a corneal transplant.

[Pathogenesis and epidemiology of neurotrophic keratopathy]. / Pathogenese und Epidemiologie der neurotrophen Keratopathie.

Neurotrophic keratopathy (NK) is a degenerative corneal disease that is based on an impairment of the corneal innervation. The damage to the sensory innervation, which is delivered through the 1st branch of the trigeminal nerve (ophthalmic nerve), can occur throughout the entire length of the nerve from the nucleus in the brainstem, e.g. caused by brain tumors, to the terminal nerve fibers in the cornea, caused for example by refractive corneal surgery (e. g. LASIK). Due to the loss of the sensory innervation, a reduced lacrimation and a reduction in the secretion of trophic factors occur. This in turn inhibits the regeneration potential of the corneal epithelium. In the most severe cases of the disease, the reduction or loss of lacrimation, together with the impaired regeneration potential of the epithelial cells, can lead to persistent epithelial defects, ulcers and corneal perforation. The NK has a prevalence of 5 or fewer individuals per 10,000 and is classified as a rare, i. e. orphan disease (ORPHA137596). A fundamental understanding of the pathogenesis and epidemiology of NK supports the early diagnosis and therefore the initiation of a specific treatment.

Correlation of central and peripheral keratometric parameters after corneal collagen cross-linking in keratoconus patients.

PURPOSE: To evaluate the difference in the central and peripheral keratometric parameters in patients with keratoconus after corneal collagen cross-linking (CXL). METHODS: Forty-eight eyes of 32 patients (18 males, 16-28 years) affected by progressive keratoconus in different stages of evolution underwent CXL using the standard epithelium-off protocol. Corneal thickness and corneal curvature before CXL and after 6 and 12 months using the Sirius tomographer were analyzed. The values of the mean corneal thickness at the corneal apex (CAT), center of the pupil (PCT), thinnest point (CTTL) and along concentric circles of 2, 4, 6, 8, 8.5, 9, 9.5 and 10 mm diameter were evaluated; the values of the mean curvature at the corneal apex and at the points in which the inferior, superior, nasal and temporal meridians crossed the above-mentioned concentric circles were also evaluated. RESULTS: The mean preoperative values for CAT, PCT and CTTL were 461.4 ± 30.3, 475.3 ± 30.5 and 441 ± 32.0, respectively. The values after 12 months of CXL were 444.6 ± 36.2, 451.6 ± 36.7 and 418.2 ± 41.4. The peripheral corneal thickness at the eight points ranged from 479 to 733 preoperatively. At 12-month post-CXL, the values ranged from 444.6 to 734.1. The mean posterior curvature from apex to periphery ranged from - 4.5 to - 9.1 days preoperatively and from - 4.5 to - 9.2 days at 12 months. These were not statistically significant (ANOVA and unpaired T test). CONCLUSIONS: Our data suggest that CXL over an 8-mm zone can stabilize the peripheral cornea. Longer-term follow-up studies on the peripheral cornea after CXL will provide useful information.

Pramlintide, an antidiabetic, is antineoplastic in colorectal cancer and synergizes with conventional chemotherapy.

BACKGROUND: Approximately 90% of patients with metastatic colorectal cancer fail therapy mainly due to resistance. Taking advantage of currently approved agents for treatment of disease conditions other than cancer for the identification of new adjuvant anticancer therapies is highly encouraged. Pramlintide is a parenteral antidiabetic agent that is currently approved for treatment of types 1 and 2 diabetes mellitus. OBJECTIVES: To address the antineoplastic potential of pramlintide in colorectal cancer and to evaluate the ability of pramlintide to enhance the cytotoxicity of 5-fluorouracil, oxaliplatin, and irinotecan against colorectal cancer cell lines expressing wild-type and mutant p53. MATERIALS AND METHODS: The antiproliferative effect of pramlintide alone or in combination with 5-fluorouracil, oxaliplatin, or irinotecan in HCT-116 and HT-29 colorectal cancer cell lines was investigated using MTT cell proliferation assay. IC50 values were calculated using Compusyn software 1.0. Synergy values (R) were calculated using the ratio of IC50 of each primary drug alone divided by combination IC50s. For each two pairs of experiments, Student's t-test was used for analysis. For combination studies, one-way analysis of variance and Tukey post hoc testing was performed using R 3.3.2 software. A p-value of <0.05 was considered significant. RESULTS: Pramlintide inhibited the growth of HCT-116 and HT-29 in a dose-dependent manner, with higher efficacy against the latter (IC50s; 48.67 and 9.10 µg/mL, respectively; p-value =0.013). Moreover, the addition of 5, 10, and 20 µg/mL of pramlintide to HCT-116 and HT-29 with 5-fluorouracil, oxaliplatin, or irinotecan induced the antiproliferative effect synergistically (R>1.6, p-value <0.05). CONCLUSION: Pramlintide enhances the cytotoxicity of conventional chemotherapy against colorectal cancer cell lines harboring wild-type or mutant p53. Thus, pramlintide is a promising potential adjuvant chemotherapy in colorectal cancer.

Air pressure changes in the creation and bursting of the type-1 big bubble in deep anterior lamellar keratoplasty: an ex vivo study.

PurposeTo measure the pressure and volume of air required to create a big bubble (BB) in simulated deep anterior lamellar keratoplasty (DALK) in donor eyes and ascertain the bursting pressure of the BB.Patients and methodsTwenty-two human sclera-corneal discs were used. Air was injected into the corneal stroma to create a BB and the pressure measured by means of a pressure converter attached to the system via a side port. A special clamp was designed to prevent air leak from the periphery of the discs. The pressure at which air emerged in the corneal tissue; the bursting pressure measured after advancing the needle into the bubble cavity and injecting more air; the volume of air required to create a BB and the volume of the BB were ascertained.ResultsType-1 BB were achieved in 19 and type-2 BB in 3 eyes. The maximum pressure reached to create a BB was 96.25+/- 21.61 kpa; the mean type-1 intrabubble pressure was 10.16 +/- 3.65 kpa. The mean bursting pressure of a type-1 BB was 66.65 +/- 18.65 kpa, while that of a type-2 BB was 14.77 +/- 2.44 kpa. The volume of air required to create a type-1 BB was 0.54 ml and the volume of a type-1 BB was consistently 0.1 ml.ConclusionsDua's layer baring DALK can withstand high intraoperative pressures compared to Descemet's membrane baring DALK. The study suggests that it could be safe to undertake procedures such as DALK-triple with a type-1 BB but not with a type-2 BB.

Vitreous cavity length in keratoconus: implications for keratoplasty.

PurposeTo compare axial length (AL) with vitreous cavity length (VCL) in patients with keratoconus and to ascertain whether graft size can be tailored to reduce myopic refractive error in patients with keratoconus undergoing penetrating keratoplasty (PK).Patients and methodsThe AL and VCL were measured prospectively in patients with keratoconus not undergoing PK (Group 1) and in normal phakic, emmetropic individuals (Group 2). A retrospective analysis of these measurements in patients with keratoconus who had undergone PK (Group 3) was also performed. The postoperative spherical equivalent (SE) was then correlated to size of donor buttons.ResultsKeratoconus patients tended to have a longer mean VCL than emmetropic normal individuals. The mean VCL of these patients (Group 1) was 16.49 mm±SD 1.13 compared to the mean VCL of 15.94 mm±SD 0.56 in normals (Group 2, P<0.0001). Patients with keratoconus who had an undersized graft showed reduced myopic refractive error compared to those with same size or oversized grafts.ConclusionVCL measurement is more accurate than AL measurement in deciding upon graft-host size disparity for corneal graft in patients with keratoconus. In patients with increased VCL, undersizing the donor button helps in reducing postoperative myopia. We recommend VCL measurement as part of the routine workup in all keratoconus patients undergoing corneal transplants.

Fibrin glue as agent for sealing corneal and conjunctival wound leaks.

PurposeTo describe a novel use of fibrin glue in managing leaking blebs and leaking wounds following trauma or surgery.MethodsInterventional case series.ResultsWe report eight patients, including three where intra-operative or immediate post-penetrating keratoplasty recalcitrant leaks from the graft-host junction and/or openings created by the needle pass, were noted. All three had thin recipient beds in the sector of leak. This was managed by intra-cameral injection of fibrin glue in the affected quadrant. This stopped the leak and allowed the defect to heal. One patient of Descemets-stripping-endothelial-keratoplasty had leak from the surgical wound, which was also sealed with fibrin glue. Two patients with leaking glaucoma-surgery-related blebs were treated with intra-bleb injection of fibrin glue to stop the leak. One patient with a penetrating corneal injury with a metal wire had a brisk leak upon removal of the wire. This was sealed with fibrin glue. Another patient of chemical burn with spontaneous leaks was managed by glue injection in the perforations. Transient rise of intraocular pressure in one patient with a leaking bleb was the only adverse event recorded.ConclusionThis novel adaptation of the application of fibrin glue can help to deal with persistent intra-operative, post-operative and traumatic aqueous and air leaks.

Corneal confocal scanning laser microscopy in patients with dry eye disease treated with topical cyclosporine.

PurposeTo investigate the effect of cyclosporine on corneal ultrastructure and on major signs and symptoms of patients with dry eye disease.Patients and methodsIn this prospective cohort study, patients with dry eye disease were treated with a drop of cyclosporine 0.05% twice daily. Clinical evaluation was carried out at baseline and at months 1, 3, and 6. All patients completed the Ocular Surface Disease Index (OSDI) questionnaire, and tear film break-up time (BUT), fluorescein and lissamine green staining, and Schirmer test were carried out. In vivo confocal microscopy was also performed and epithelial cellular density, keratocyte activation, and subbasal plexus morphology were assessed.ResultsA total of 40 patients completed the study. After 6 months, OSDI, BUT, and fluorescein and lissamine green staining showed a clinically significant improvement. During the 6-month follow-up, density of intermediate epithelial cells increased from 1969.5±85.4 cell/mm2 to 4881.2±175.7 cell/mm2 (P<0.01); average grade of keratocyte activation decreased from 3.6±0.1 to 1.8±0.1 (P<0.001); average grade of number of subbasal nerves decreased from 5.3±0.2 to 2.6±0.2 (P<0.001); average grade of nerve reflectivity decreased from 3.8±0.1 to 2.1±0.2 (P<0.001); and average grade of nerve tortuosity decreased from 3.8± 0.1 to 2.2±0.2 (P<0.001).ConclusionCyclosporine was effective in controlling symptoms and signs of dry eye disease. In vivo confocal microscopy showed an increase in cell density of intermediate epithelium cells, a decrease in hyperreflective keratocytes, and a decrease in density, tortuosity, and reflectivity of nerve fibers.

Pre-Descemets endothelial keratoplasty: the PDEK clamp for successful PDEK.

PurposeTo design and produce a clamp that enables good handling of donor sclera-corneal disks, allows air to be injected in the corneal stroma and consistently provides pre-Descemets endothelial keratoplasty (PDEK) tissue by creation of a type-1 big bubble (BB) avoiding a type-2BB by shutting the fenestrations in the periphery of the pre-Descemets layer and preventing escape of air.Patients and methodsA clamp with spring loaded handles attached to two rings of 9 mm diameter and 1 mm width with a side port for insertion of a needle for air injection was designed and produced. The clamp was tested in 20 human donor sclera-corneal disks and refined over three prototypes. The final design was used for PDEK in 8 patients.ResultsThe clamp was able to prevent any escape of air or the formation of a type-2BB. By preventing air leak, it provided better control over the volume of air injected allowing for consistent formation of a type-1BB, which could be expanded to its full diameter by advancing the tip of the needle into the bubble cavity. Centration of tissue in the clamp was important. The clamp was used successfully to obtain PDEK tissue from 8 donor sclara-corneal disks for transplant in three eyes with pseudophakic bullous keratopathy and five eyes with pseudophakia and Fuchs' endothelial dystrophy.ConclusionThe PDEK clamp is a simple and useful instrument that will give surgeons the confidence of consistently obtaining PDEK tissue with ease and without the risk of separating the Descemets membrane in a type-2BB.

Optical coherence tomography characteristics of different types of big bubbles seen in deep anterior lamellar keratoplasty by the big bubble technique.

PurposeTo define optical coherence tomography (OCT) characteristics of type-1, type-2, and mixed big bubbles (BB) seen in deep anterior lamellar keratoplasty.MethodsHuman sclero-corneal discs were obtained from UK (30) and Canada (16) eye banks. Air was injected into corneal stroma until a BB formed. UK samples were fixed in formalin before scanning with Fourier-domain (FD-OCT). One pair of each type of BB was scanned fresh. All BB obtained from Canada were scanned fresh with time-domain (TD-OCT). For each OCT machine used, type-1 BB from which Descemets membrane (DM) was partially peeled, were also scanned. The morphological characteristics of the scans were studied.ResultsFD-OCT of the posterior wall of type-1 (Dua's layer (DL) with DM) and type-2 BB (DM alone) both revealed a double-contour hyper-reflective curvilinear image with a hypo-reflective zone in between. The anterior line of type-2 BB was thinner than that seen with type-1 BB. In mixed BB, FD-OCT showed two separate curvilinear images. The anterior image was a single hyper-reflective line (DL), whereas the posterior image, representing the posterior wall of type-2 BB (DM) was made of two hyper-reflective lines with a dark space in between. TD-OCT images were similar with less defined component lines, but the entire extent of the BB could be visualised.ConclusionOn OCT examination the DM and DL present distinct features, which can help identify type-1, type-2, and mixed BB. These characteristics will help corneal surgeons interpret intraoperative OCT during lamellar corneal surgery.

Fibrin glue inhibits migration of ocular surface epithelial cells.

PurposeFibrin glue has been used successfully in numerous ophthalmic surgical procedures. Recently, fibrin glue has been used in limbal stem cell transplantation to reduce both operative time and to negate the need for sutures. The aim of this study was to determine the effects of fibrin glue on epithelial cell migration in vitro.MethodsCorneoscleral rims were split to retain the epithelial layer, Bowman's layer, and anterior stroma. Rims were cut into eight equal-sized pieces and were placed directly on culture plates or affixed with fibrin glue. Rims were maintained in culture for 25 days and epithelial cell growth was monitored. Cells were photographed to measure area or growth and immunofluorescence staining of explants for fibrin was performed.ResultsExplants that were glued demonstrated significantly delayed epithelial cell growth and migration as compared with explants without glue. By day 16, all fibrin glue had dissolved and coincided with onset of cell growth from glued explants. Cell growth commenced between days 3 and 4 for control explants without glue and around days 14-16 for explants with fibrin glue.ConclusionsFibrin glue delays epithelial cell migration by acting as a physical barrier and can potentially interfere with explant-derived limbal epithelial cell migration on to the corneal surface. We propose that glue should be used to attach the conjunctival frill of the limbal explant but care should be taken to ensure that the glue does not wrap around the explant if used to secure the explant as well. Strategic use of glue, to attach the recessed conjunctiva, can be advantageous in delaying conjunctival cell migration and reducing the need for sequential sector conjunctival epitheliectomy.

Deep anterior lamellar keratoplasty--triple procedure: a useful clinical application of the pre-Descemet's layer (Dua's layer).

PURPOSE: To describe a technique which exploits the transparency and toughness of the pre-Descemets layer (Dua's layer) to safely perform deep anterior lamellar keratoplasty (DALK) and phacoemulsification at the same time. METHODS: Three DALK procedures combined with phacoemulsification were performed by the same surgeon using the big-bubble (BB) technique at the Research Institute of Ophthalmology, Cairo, Egypt. In two cases a type-1 BB, baring Dua's layer, was achieved and in one case a type-2 BB, baring the Descemet's membrane (DM), was achieved. The surgeries were video recorded and photo slit-lamp images were taken in the follow-up visits. RESULTS: DALK with phacoemulsification and lens implant was carried out in both patients where a type-1 BB was achieved. At a follow up of 18 months (first case) and 6 months (second case), the best corrected vision was 6/12 for each. In the case where a type-2 BB was achieved, the DM ruptured during injection of viscoelastic prior to capsulorhexis. The procedure was converted to a penetrating keratoplasty without phacoemulsification. CONCLUSION: When a type-1 BB is achieved simultaneous DALK and phacoemulsification can be safely accomplished. Dua's layer allows a clear view for performing phacoemulsification with the added benefit of its toughness, which can maintain a stable anterior chamber for cataract surgery. This should not be attempted when a type-2 BB is achieved.

Sequential selective same-day suture removal in the management of post-keratoplasty astigmatism.

AIMS: In a previous study, we proposed that corneal topography performed 30-40 min after the initial suture removal can identify the next set of sutures requiring removal, for the treatment of post-keratoplasty astigmatism. The aim of this study was to evaluate the effect of removing subsequent sets of sutures at the same sitting. METHODS: 10/0 nylon interrupted sutures were placed, to secure the graft-host junction, at the time of keratoplasty. Topography was performed using Pentacam (Oculus) before suture removal. The sutures to be removed in the steep semi-meridians were identified and removed at the slit-lamp biomicroscope. Topography was repeated 30-40 min post suture removal, the new steep semi-meridians determined, and the next set of sutures to be removed were identified and removed accordingly. Topography was repeated 4-6 weeks later and the magnitude of topographic astigmatism was recorded. A paired-samples t-test was used to evaluate the impact of selective suture removal on reducing the magnitude of topographic and refractive astigmatism. RESULTS: Twenty eyes of 20 patients underwent sequential selective same-day suture removal (SSSS) after corneal transplantation. This study showed that the topographic astigmatism decreased by about 46.7% (3.68 D) and the refractive astigmatism decreased by about 37.7% (2.61 D) following SSSS. Vector calculations also show a significant reduction of both topographic and refractive astigmatism (P<0.001). CONCLUSION: SSSS may help patients to achieve satisfactory vision more quickly and reduce the number of follow-up visits required post keratoplasty.

Intra-lesional 5 fluorouracil for the management of recurrent pterygium.

AIM: Recurrence is the most common complication arising from pterygium surgery. The aim of this study was to investigate the effectiveness of 5 fluorouracil (5FU) in halting the recurrence of pterygium after surgical excision. METHODS: A retrospective review of patients treated for pterygium recurrence was carried out. Patients with recurrent (secondary) pterygium were treated with multiple weekly intra-lesional injections of 0.1-0.2 ml (2.5-5 mg) 5FU post-operatively depending on the size of the recurrence. The treatment was started within 1 month from the date of recurrence. The time from surgery to start of recurrence, previous treatment modalities, and number of recurrences were documented. The number of injections required to induce arrest of progression and/or regression of vascularity and fleshiness of the pterygium and any complications related to 5FU treatment were examined. RESULTS: Fifteen eyes from 14 patients with recurrent pterygium treated with intra-lesional 5FU injections were analysed. Three of the 15 eyes had undergone a secondary excision and 12 had undergone a primary excision. In all, 93.3% of patients showed regression of the fibrovascular tissue (thickness and vascularity) and arrest of progression following a dose of 0.1-0.2 ml (2.5-5 mg) 5FU. Twelve eyes required three injections or fewer, whereas one patient required eight injections. This beneficial effect was maintained over an average follow-up period of 17 months. No complications from 5FU were observed. CONCLUSION: The use of weekly intra-lesional 5FU injections for the treatment of recurrent pterygium is safe and effective in limiting the progression and inducing the regression of recurrent pterygium. The number of injections can be tailored according to clinical need.

Antimicrobial peptides expression by ocular surface cells in response to Acanthamoeba castellanii: an in vitro study.

AIMS: Antimicrobial peptides (AMPs) are natural effectors of the innate immune response. Much work has been done to study their response and effects on bacterial and viral infection. Little if any information is available in relation to protozoal infections. The aim of the study was to comprehensively study the gene expression of the ocular AMPs in human corneal limbal epithelial cells stimulated with Acanthamoeba castellanii (AC). METHODS: Human corneal limbal epithelial cells were exposed to AC at different time points, up to 9 h, the genomic profile of the AMPs were analysed at these time point using real time PCR. corneal limbal epithelial cells not infected with AC were used as controls. RESULTS: Seven of the eight studied AMPs showed statistically significant upregulation in gene expression. Human beta Defensin 3 (hBD3) showed a very significant 10-fold upregulation in the exposed cells and Ribonuclease-7 (RNase-7) showed a very early and consistent increase. Human beta Defensin 1 (hBD1) was the only downregulated AMP. CONCLUSIONS: The study data suggests a possible role of the AMPs in combating the amoebic infection at the ocular surface. Using AMPs singly or in combination is a promising avenue for further exploration in the treatment of the sight threatening Acanthamoeba keratitis.

Emerging treatment paradigms of ocular surface disease: proceedings of the Ocular Surface Workshop.

The objective of the Ocular Surface Workshop in Rome, Italy, on 6 February 2009, was to enhance the understanding of ocular surface disease (OSD) through an exploration of the nature of its complexities and current treatment paradigms across Europe. It was hoped that the peer-to-peer discussions and updates regarding common knowledge, clinical practices and shared experiences at this workshop would subsequently shape future treatment approaches to OSD.