The aim of the present paper is to describe the morphological changes that occur in human corneal endothelium as an immediate consequence of corneal cryopreservation. Therefore, 16 human donor corneas were cryopreserved with an original procedure at a 1 °C/min cooling rate in a freezing solution cryoprotected with 7% dimethylsulphoxide until a final temperature of –100 °C was reached. After storage of the corneas in liquid nitrogen for periods ranging from 1 to 96 days (mean: 34.31 days), the corneas were thawed in a water bath at +37 °C. Eight additional control corneas were processed without cryopreservation. Morphological assessment of the endothelial layer was performed by scanning electron microscopy and trypan blue and alizarin red S vital staining. Results showed cryoinduced damage at variable degrees in all cryopreserved corneas. They were classified into three groups according to the intensity and extension of the cryoinduced damage: group I (n = 10): corneas with minor endothelial alterations consisting in the presence of microholes in the posterior cell membrane; group II (n = 1): corneas with generalized disruption of endothelial intercellular junctions and intact cell membranes; group III (n = 5): corneas with severe endothelial damage consisting of massive cell necrosis and complete alteration of the morphological pattern of the endothelium. All control corneas had intact endothelial layers. Cryoinduced damage cannot be completely avoided with the cryopreservation protocol tested. The high interindividual variability of the results observed is not related to the storage time of the cornea in liquid nitrogen.

In the present study eight human eyeballs were specifically prepared for scanning-electron-microscopic observation of the zonule. The zonule consisted of two main layers of radial fibres, an anterior and a posterior one, that inserted on the anterior and the posterior lens capsules, respectively. Some fibres inserted on the equator of the lens. Posterior zonular fibres originated at the pars plana, entered the dorsal part of the ciliary valleys and then changed their direction towards the posterior face of the lens. Posterior fibres inserted on the posterior capsule of the lens by branched endings 1 mm behind the equator of the lens. Anterior zonular fibres originated mainly at the pars plana and occasionally at the ciliary valleys. After running completely through the ciliary valleys in close contact with the lateral walls of the ciliary processes, they changed their direction at the anterior endings of the pars plicata and reached the anterior lens capsule. Anterior zonular insertions were achieved by webbed endings that diffused into the anterior capsule 2 mm in front of the lens equator. The extraordinary distension capacity of the zonular fibres was demonstrated by pulling the anterior lens capsule after hydrodissection. As a consequence, the anterior fibres were stretched up to four times their original length without breaking or disinserting.

The purpose of the present study was to set up and test a cryopreservation method for long-term storage of human corneas. Therefore the freezing solution was optimized in 264 rabbit corneas by testing the type of cryoprotectant, its concentration, addition and dilution pattern and exposure temperature. Then rabbit corneas were frozen in the optimum solution at different cooling rates and thawed in a water bath at different temperatures. Eight human corneas were cryopreserved with the method showing optimum results in rabbit corneas and four additional corneas were used as controls. Endothelial viability was assessed after each step by vital staining and scanning electron microscopy. Best results after exposure of rabbit corneas to the freezing solution were achieved when using a 10% cryoprotectant concentration, with direct addition/dilution and exposure at room temperature (3512 +/-300 viable cells mm(2) when using dimethylsulfoxide; 3403 +/- 245 viable cells mm(2) when using 1,2-propanediol). Cryopreserved rabbit corneas had the highest endothelial cell survival when frozen at 1 degrees C/min and thawed at 37 degrees C (2003 +/- 372 viable cells/mm(2) when using dimethylsulfoxide and 1357 +/- 667 viable cells/mm(2) when using 1,2-propanediol). Cryopreserved human corneas had 753 +/- 542 viable cells/mm(2) when using dimethylsulfoxide and 56 +/- 56 viable cells/mm(2) when using 1,2-propanediol. We can conclude that the method developed is easy to handle and shows optimum results in rabbit corneas, with an endothelial cell survival that is consistent with transplant acceptability criteria. The results obtained in human corneas are below prediction and are still unsatisfactory for successful use in eye banking.

PURPOSE: To compare the morphological features of photorefractive ablations produced by six different excimer lasers. METHODS: A spherocylindrical photoablation (-2.00 -2.00 x 90°; 6-mm optical zone) was performed on regular fluence plates with six excimer lasers: Bausch & Lomb Technolas 217C, Schwind Esiris, Kera Technology Isobeam D200, Ladarvision 4000, Zeiss Meditec MEL 70 G-Scan, and Visx Star S3. Morphometric analysis of the fluence plates provided superficial measurements of the ablated areas. RESULTS: Two areas were identified visually: a central area with a complete ablation of the metallic surface layer and a surrounding area with a partial ablation. The dimensions of the ablated areas were highly variable in the lasers tested. The major differences appeared in the total ablated area (ranging from 38.55 mm^sup 2 ^ [Schwind] to 81.94 mm p 2 [Bausch & Lomb] and in the peripheral to total area ratio (ranging from 36.95% [Schwind] to 59.51% [Ladarvision]). CONCLUSION: Large differences appeared in the superficial dimensions and contours of the ablations produced by different excimer lasers for the treatment tested in this study. It remains unknown how these different ablation patterns induce the same optical correction, but we assume that the depth of the ablation compensates for the differences in the surface extension of the ablated areas. [J Refract Surg 2004;20:106-109]

By studying the eyes of 20 human fetuses 33–326 mm in length (8 to 36 weeks gestation) with the light and scanning electron microscope we show that the sclerocorneal and the uveal trabecula cannot be considered as independent structures, since embryologically and morphologically they are the sclero-corneo-iridal insertions of the longitudinal fibres of the ciliary muscle.