RESUMEN
PURPOSE: Different molecular targets, such as the epidermal growth factor receptor, have been identified for the prophylaxis of posterior capsule opacification. This led to the proposal of several drugs, yet drug delivery into the capsular bag remains challenging. The intraocular lens as a drug delivery device would provide a convenient method to allow drug release in the location needed. This is to evaluate the effect of a drug-eluting intraocular lens using an epidermal growth factor receptor inhibitor. METHODS: Hydrophobic and hydrophilic intraocular lenses were coated with gefitinib using the dip coating technique. The cellular response on the modified intraocular lenses was tested in a human lens epithelial cell line (FHL-124) in an anterior segment model. Furthermore, modified intraocular lenses were implanted into human capsular bags ex vivo. Drug release was determined as well as the biocompatibility on human corneal endothelial cells. Unmodified intraocular lenses served as controls. In addition, immunofluorescence staining with fibronectin as a marker for fibrotic response was conducted. RESULTS: Both coated hydrophilic and hydrophobic intraocular lenses could attenuate the cell growth of FHL-124 cells in the human capsular bag in comparison to the unmodified controls. Furthermore, gefitinib-soaked intraocular lenses showed a constant drug release over the first 10 days. No reduction in cell viability of corneal endothelial cells occurred. A decrease in fibronectin expression under gefitinib treatment could be observed. CONCLUSION: In vitro epidermal growth factor receptor seems to be a valuable target for the prevention of posterior capsule opacification. The gefitinib-eluting intraocular lens in this study could inhibit cell growth in non-toxic concentrations.
Asunto(s)
Opacificación Capsular/prevención & control , Portadores de Fármacos , Receptores ErbB/antagonistas & inhibidores , Gefitinib/administración & dosificación , Lentes Intraoculares , Inhibidores de Proteínas Quinasas/administración & dosificación , Línea Celular , Proliferación Celular/efectos de los fármacos , Sistemas de Liberación de Medicamentos , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Fibronectinas/metabolismo , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Ensayo de Materiales , Cápsula Posterior del Cristalino/efectos de los fármacosRESUMEN
BACKGROUND: Posterior capsule opacification (PCO) after cataract surgery is influenced by intraocular lens (IOL) design and material. The following is an ex vivo comparison of PCO between the Clareon vs. the AcrySof IOL in human capsular bags. METHODS: Twenty cadaver capsular bags from 10 human donors were used, with the novel hydrophobic IOL (Clareon, CNA0T0) being implanted in one eye and the other eye of the same donor receiving the AcrySof IOL (SN60WF) following phacoemulsification cataract surgery. Five capsular bags of 3 donors served as controls without IOL. Cellular growth of lens epithelial cells was photo-documented daily. The primary endpoint was the time until full coverage of the posterior capsule by cells. Furthermore, immunofluorescence staining of capsular bags for the fibrotic markers f-actin, fibronectin, alpha smooth muscle actin, and collagen type 1 were performed. RESULTS: The new Clareon IOL did not show any disadvantages in terms of days until full cell coverage of the posterior capsule in comparison to the AcrySof (p > 0.99). Both, the Clareon (p = 0.01, 14.8 days) and the AcrySof IOL (p = 0.005, 15.7 days) showed a slower PCO development in comparison to the control (8.6 days). The fibrotic markers f-actin, fibronectin, alpha smooth muscle actin, and collagen type 1 were equally distributed between the two IOLs and differed from the control. CONCLUSIONS: A comparable performance has been found in the ex vivo formation of PCO between the two IOLs. Long-term clinical studies are necessary to reach final conclusions.
Asunto(s)
Opacificación Capsular/diagnóstico , Implantación de Lentes Intraoculares , Lentes Intraoculares , Facoemulsificación , Cápsula Posterior del Cristalino/patología , Actinas/metabolismo , Anciano , Opacificación Capsular/metabolismo , Células Cultivadas , Colágeno Tipo I/metabolismo , Fibronectinas/metabolismo , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , Persona de Mediana Edad , Cápsula Posterior del Cristalino/metabolismo , Diseño de Prótesis , Donantes de Tejidos , Agudeza Visual/fisiologíaRESUMEN
PURPOSE: To determine whether excimer laser ablation of guttae is a viable strategy for removal of diseased tissue in Fuchs' endothelial corneal dystrophy (FECD) on excised human Descemet membranes and whether an excimer laser-created wound on healthy human corneas ex vivo is recolonized with corneal endothelial cells. METHODS: Descemet membranes of FECD patients and corneal endothelium of normal human corneas were ablated ex vivo using an excimer laser licensed for glaucoma surgery. Specimens were kept in cell culture medium supplemented with 10 µm of rho-kinase inhibitor ripasudil. Corneal endothelial cell regeneration was observed using light and electron scanning microscopy. Furthermore, the whole corneal samples were evaluated by haematoxylin/eosin staining and immunohistochemical analysis using antibodies against Na+ /K+ -ATPase. RESULTS: Guttae and corneal endothelium could be ablated with an excimer laser without total ultrastructural damage to the Descemet membrane or stroma. Nearly complete endothelial wound closure was accomplished after 26-38 days in treated corneas. Light and electron scanning microscopy suggested the establishment of a layer of flat endothelial cells. Additionally, Na+ /K+ -ATPase expression could only be observed on the inner side of the Descemet membrane. CONCLUSION: Our proof of concept study demonstrated that excimer lasers can be used to ablate diseased tissue from excised FECD Descemet membranes ex vivo. Additionally, corneal endothelial cells recolonize a previously ablated endothelial area in healthy human corneas ex vivo under treatment with ripasudil. Thus, our results are the first experimental basis to further investigate the feasibility of an excimer laser ablation as a graftless FECD treatment option.
Asunto(s)
Lámina Limitante Posterior/cirugía , Endotelio Corneal/cirugía , Distrofia Endotelial de Fuchs/cirugía , Catarata , Córnea/cirugía , Trasplante de Córnea/métodos , Lámina Limitante Posterior/ultraestructura , Endotelio Corneal/ultraestructura , Humanos , Terapia por Láser/métodos , Láseres de Excímeros , Prueba de Estudio ConceptualRESUMEN
PURPOSE: To compare the effect of different laser pulse energy settings in femtosecond laser-assisted cataract surgery with that of standard phacoemulsification and no energy at all used on posterior capsule opacification (PCO) in vitro. SETTING: Cell and Molecular Biology Research Laboratory, Department of Ophthalmology, Ludwig-Maximilians-University Munich, Real Eyes, Ophthalmology Center, Munich, and Institute for Clinical Pathology, Goethe University Frankfurt, Frankfurt, Germany. DESIGN: Experimental study. METHODS: Fifteen cadaver capsular bags were cultivated from 8 human donors under standard cell culture conditions. For preparation of the capsular bag, 4 groups were established as follows: femtosecond laser-assisted cataract surgery standard energy (n = 3), femtosecond laser-assisted cataract surgery high energy (n = 3), phacoemulsification (n = 6), and hydrodissection without energy (extracapsular cataract extraction) (n = 3). Growth of lens epithelial cells was observed and photodocumented. The days until full cell coverage of the posterior capsule were documented. Capsular bags were stained for fibronectin, α-smooth muscle actin, and collagen type 1. RESULTS: Cell growth patterns in all treatment groups were comparable, with no statistically significant differences detected at any timepoint measured (P = .81, Kruskal-Wallis). The markers for fibrosis were equally distributed in all groups, indicating an equal fibrotic reaction in all groups. CONCLUSION: Femtosecond laser-assisted cataract surgery did not increase different cellular responses in PCO formation comparison with phacoemulsification in vitro, even when higher laser pulse energy levels were used.
Asunto(s)
Opacificación Capsular/etiología , Terapia por Láser/efectos adversos , Cápsula del Cristalino/patología , Facoemulsificación/efectos adversos , Complicaciones Posoperatorias , Anciano , Anciano de 80 o más Años , Cadáver , Opacificación Capsular/patología , Extracción de Catarata/efectos adversos , Fibrosis/etiología , Fibrosis/patología , Humanos , Cápsula del Cristalino/cirugía , Persona de Mediana EdadRESUMEN
PURPOSE: Posterior capsule opacification (PCO) still represents the main long-term complication of cataract surgery. Research into pharmacologic PCO prophylaxis is extensive. One promising candidate drug is methotrexate (MTX). Our aim is to determine the in vitro feasibility of MTX-loaded poly(lactic-co-glycolic) (PLGA) biomatrices sprayed on intraocular lenses (IOLs) as a drug-delivery implant. METHODS: Hydrophilic and hydrophobic acrylic IOLs were spray-coated with MTX-loaded PLGA. Unsprayed, solvent only, and solvent-PLGA-sprayed IOLs served as controls. All IOLs were evaluated for their growth-inhibiting properties in an in vitro anterior segment model and the ex vivo human capsular bag. The release kinetics of MTX from the IOLs was determined. The toxicity of MTX on corneal endothelial cells was evaluated by using a dye reduction colorimetric assay. MTX was also used in a scratch assay. RESULTS: MTX-PLGA-IOL showed a significant difference in cell proliferation and migration compared with all controls in the anterior segment model (p < 0.001) and in the human capsular bag model (p = 0.04). No difference in viability was observed on corneal endothelial cells (p = 0.43; p = 0.61). MTX significantly inhibited cells in the scratch assay (p = 0.02). At all measured points, the released MTX dose remained above EC50 and below the toxic dose for the endothelium. CONCLUSIONS: In view of the strong inhibition of PCO in vitro with the lack of toxic effects on a corneal cell line, MTX encapsulating microspheres seem to be a promising method for modifying IOL.