Primary lens instability in ten related cats: clinical and genetic considerations.

OBJECTIVES: To describe bilateral lens instability in 10 related domestic shorthair cats over three generations. METHODS: Complete ophthalmic examinations were performed. Lentectomies were carried out. Sections of affected lenses focused on the equatorial area were examined by transmission electron microscopy. The potential involvement of several candidate genes (ADAMTS17, ADAMTSL4, ADAMTS10 and FBN1) known to be associated with lens luxation in other species was investigated. RESULTS: The group of animals included 10 related cats, nine of them being affected by lens instability over three generations. Transmission electron microscopy showed the presence of zonular material at the lens equator. Signs of lens instability were not associated with other ocular disease. Analysis of the pedigree suggests a dominantly inherited condition. A mutation in ADAMTS17 was excluded, but a possible association between the condition and a microsatellite flanking FBN1 indicates this gene should be considered a strong candidate responsible for primary lens luxation in this pedigree. CLINICAL SIGNIFICANCE: These observations suggest an inherent zonular defect unrelated to extraneous factors. The family relationship is compatible with a possible genetic basis, and the pedigree suggests that the condition could be dominant. Data also suggest the mutation in the FBN1 gene could be responsible for primary lens luxation in this pedigree of cats.

The outer limiting membrane (OLM) revisited: clinical implications.

PURPOSE: The outer limiting membrane (OLM) is considered to play a role in maintaining the structure of the retina through mechanical strength. However, the observation of junction proteins located at the OLM and its barrier permeability properties may suggest that the OLM may be part of the retinal barrier. MATERIAL AND METHODS: Normal and diabetic rat, monkey, and human retinas were used to analyze junction proteins at the OLM. Proteome analyses were performed using immunohistochemistry on sections and flat-mounted retinas and western blotting on protein extracts obtained from laser microdissection of the photoreceptor layers. Semi-thin and ultrastructure analyses were also reported. RESULTS: In the rat retina, in the subapical region zonula occludens-1 (ZO-1), junction adhesion molecule (JAM), an atypical protein kinase C, is present and the OLM shows dense labeling of occludin, JAM, and ZO-1. The presence of occludin has been confirmed using western blot analysis of the microdissected OLM region. In diabetic rats, occludin expression is decreased and glial cells junctions are dissociated. In the monkey retina, occludin, JAM, and ZO-1 are also found in the OLM. Junction proteins have a specific distribution around cone photoreceptors and Müller glia. Ultrastructural analyses suggest that structures like tight junctions may exist between retinal glial Müller cells and photoreceptors. CONCLUSIONS: In the OLM, heterotypic junctions contain proteins from both adherent and tight junctions. Their structure suggests that tight junctions may exist in the OLM. Occludin is present in the OLM of the rat and monkey retina and it is decreased in diabetes. The OLM should be considered as part of the retinal barrier that can be disrupted in pathological conditions contributing to fluid accumulation in the macula.

Effect of procyanidolic oligomers on corneal collagen of rabbits treated by excimer laser photoablation.

Procyanidolic oligomers (PCO) are mainly used for their therapeutic effect on the vascular wall. We could show that the mechanism of this effect involves interactions with mesenchymal cells and extracellular matrix (ECM). Recently we demonstrated in vitro that they also act on cornea, a tissue rich in ECM. For instance they stimulate the corneal biosynthesis of type VI collagen and proteoglycans. A potent antiprotease effect also could be demonstrated on corneas. In our present work we examined in vivo action of PCO on corneas. A group of rabbits received during 1 week on their right eye a treatment by procinaidolic oligomers, the left eyes were kept as controls. After 1 week both eyes underwent excimer laser photobalation. Another group of rabbits also received the PCO treatment on the right eye, not on the left, but was not treated by photoablation. One week after the surgical intervention corneas were collected and biochemical and morphological observations were carried out. Ocular administration of PCO was well tolerated and no toxic or inflammatory side-effects could be seen. In the control group photoablation was followed by a decrease of the content of corneas in type I collagen and a strong increase in type III collagen. On the corneas treated by PCO these alterations of the composition were not observed. These results indicate that PCO treatment before excimer laser photoablation maintains within normal limits the biochemical composition of the operated corneas.

Treatment of ocular cicatricial pemphigoid with sulfasalazine.

PURPOSE: To assess the outcome of patients with ocular cicatricial pemphigoid (OCP) treated with sulfasalazine as an alternative to dapsone. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Nine patients with biopsy-proven OCP and previous dapsone-related adverse effects (hemolysis and gastrointestinal disturbances) treated with oral sulfasalazine. METHODS: Clinical data were abstracted from patients' medical records. MAIN OUTCOME MEASURES: Patients' symptoms, ocular inflammation, conjunctival scarring, complete blood cell count (including reticulocyte count). RESULTS: At the initiation of sulfasalazine therapy, ocular inflammation was controlled in all patients but one. Mean follow-up was 12 months (range, 2-35 months). Median oral sulfasalazine dosage was 3 g (range, 1-4 g). The disease remained controlled with sulfasalazine alone in four patients (45%). Two patients (22%) required adjunctive oral cyclophosphamide. Adverse effects necessitating drug withdrawal occurred in three patients (33%): hemolysis in two and gastrointestinal disturbances in one. CONCLUSIONS: Sulfasalazine may be useful in OCP patients with previous dapsone-related adverse effects.

Chemotactic penetration of keratocytes in ePTFE polymer in vitro.

Expanded polytetrafluoroethylene (ePTFE) is used as a support for artificial corneas. Implanted in corneas, most of the time this polymer is colonized by corneal host cells. The absence of colonization often coincides with extrusion of the polymer. Therefore, we decided to introduce keratocytes into ePTFE in vitro before implantation. Because keratocytes do not spontaneously enter ePTFE, we used several chemoattractants, separately and in a mixture, to stimulate the penetration of cultured keratocytes into the polymer. The influence of the passage number on cell penetration was also studied. No significant differences were observed up to the seventh passage, although seventh-passage cells penetrated somewhat more slowly than younger cells. Satisfactory results were obtained with four of the tested chemotactic factors: IL-6, type alpha transforming growth factor (TGF-alpha), platelet derived growth factor isoform BB (PDGF-BB), and fibroblast growth factor-2 (FGF-2). Under our experimental conditions, two to more than six million keratocytes entered the polymer discs with a volume of 706.5 mm(3) in the presence of these four chemoattractants. TGF-alpha was the most efficient and was selected for further in vitro and in vivo studies.

Histologic phenotype-genotype correlation of corneal dystrophies associated with eight distinct mutations in the TGFBI gene.

PURPOSE: To establish a phenotype-genotype correlation of various autosomal-dominant corneal dystrophies among French subjects. DESIGN: Retrospective molecular genetic study and clinicopathologic correlation. PARTICIPANTS: Forty-four subjects from 26 unrelated French families were included in this study, and 60 corneal buttons could be examined at the histologic and ultrastructural levels. METHODS: Light microscopy and transmission electron microscopy were performed on corneal specimens obtained during keratoplasty. Blood samples were collected for DNA analysis. MAIN OUTCOME MEASURES: After genomic DNA extraction from peripheral blood leukocytes of each family member, exons of the TGFBI gene were amplified by polymerase chain reaction (PCR), and the PCR products were directly sequenced on both strands. RESULTS: Four different mutations were found to be responsible for dystrophy of granular type (R555W, R124L, R124H, and R124L+delT125-delE126), three other different mutations produced a lattice type (R124C, H626R, and A546T), and the last mutation identified was associated with the honeycomb-shaped dystrophy (R555Q). Each subtype of dystrophy showed, histologically and ultrastructurally, specific characteristics that are easily recognizable. However, besides these stereotyped forms, differential histologic diagnosis of atypical forms remains difficult, and these forms could be misdiagnosed. CONCLUSIONS: The characteristic biomicroscopic appearance and histopathologic features of each "classic" dystrophy present a significant degree of specificity and generally provide an accurate diagnosis. However, atypical forms in which clinical and histologic data alone could be misleading, are unequivocally diagnosed after DNA analysis.

EIU in the rat promotes the potential of syngeneic retinal cells injected into the vitreous cavity to induce PVR.

PURPOSE: To determine whether syngeneic retinal cells injected in the vitreous cavity of the rat are able to initiate a proliferative process and whether the ocular inflammation induced in rats by lipopolysaccharide (LPS) promotes this proliferative vitreoretinopathy (PVR). METHODS: Primary cultured differentiated retinal Müller glial (RMG) and retinal pigmented epithelial (RPE) cells isolated from 8 to 12 postnatal Lewis rats were injected into the vitreous cavity of 8- to 10-week-old Lewis rats (10(5) cells/eye in 2 microlieter sterile saline), with or without the systemic injection of 150 microgram LPS to cause endotoxin-induced uveitis (EIU). Control groups received an intravitreal injection of 2 microliter saline. At 5, 15, and 28 days after cell injections, PVR was clinically quantified, and immunohistochemistry for OX42, ED1, vimentin (VIM), glial fibrillary acidic protein (GFAP), and cytokeratin was performed. RESULTS: The injection of RMG cells, alone or in combination with RPE cells, induced the preretinal proliferation of a GFAP-positive tissue, that was enhanced by the systemic injection of LPS. Indeed, when EIU was induced at the time of RMG cell injection into the vitreous cavity, the proliferation led to retinal folds and localized tractional detachments. In contrast, PVR enhanced the infiltration of inflammatory cells in the anterior segment of the eye. CONCLUSIONS: In the rat, syngeneic retinal cells of glial origin induce PVR that is enhanced by the coinduction of EIU. In return, vitreoretinal glial proliferation enhanced the intensity and duration of EIU.

[Avellino dystrophy. Current diagnostic criteria]. / La dystrophie d'Avellino. Critères diagnostiques modernes.

We report a French family suffering from an Avellino corneal dystrophy diagnosed by using clinical, histological, ultrastructural and genetics findings. Our results indicate that direct corneal examination and routine histological examinations must always be associated with an assay for BIGH3 gene mutations to establish a modern and unambiguous diagnosis of a corneal dystrophy.

Clinical, histologic, and ultrastructural features of the corneal dystrophy caused by the R124L mutation of the BIGH3 gene.

OBJECTIVE: This study was designed to describe the clinical, histologic, and ultrastructural features of the corneal dystrophy associated with the R124L mutation of the BIGH3 gene. DESIGN: Retrospective clinical and histologic review of a new genetic mutation. PARTICIPANTS: Thirty-four patients from five unrelated French families with corneal dystrophy caused by the R124L mutation of the BIGH3 gene were studied at the clinical, histologic, and ultrastructural levels. Records of patients carrying this mutation were compared with those from three unrelated patients with corneal dystrophy of Bowman's layer (CDB) type 2 (R555Q mutation) and from three unrelated patients with classic corneal granular dystrophy (R555W mutation). INTERVENTION: The mutational genetic status of the BIGH3 gene was determined for each patient, and the histologic and ultrastructural data available after corneal graft were analyzed. MAIN OUTCOMES MEASURES: Genomic DNA was extracted from peripheral blood leukocytes. Exons 4 and 12 of the BIGH3 gene were amplified by the polymerase chain reaction (PCR), and the PCR products were directly sequenced. RESULTS: All 34 patients with the R124L mutation displayed the clinical, histologic, and electron microscopic features of the dystrophy previously described as a superficial variant of corneal granular dystrophy. Combining molecular genetics with clinical and histologic findings established a clear distinction between the R555Q and R555W dystrophies. CONCLUSIONS: The R124L mutation of the BIGH3 gene is associated with specific clinical and morphologic criteria. This indicates that molecular studies are needed for an adequate classification of corneal dystrophies. All criteria are presently available to segregate the dystrophy caused by the R124L mutation (known as CDB1) from the dystrophy caused by the R555Q mutation (known as CDB2).

A viscous bioerodible poly(ortho ester) as a new biomaterial for intraocular application.

The biocompatibility of a viscous, hydrophobic, bioerodible poly(ortho ester) (POE) intended for intraocular application was investigated. POE was evaluated as a blank carrier and as containing modulators of degradation. Each formulation was injected intracamerally and intravitreally in rabbit eyes, and clinical and histological examinations were performed postoperatively for 2 weeks. In the case of intracameral injections, polymer biocompatibility appeared to depend on the amount injected in the anterior chamber. When 50 microL was administered, the polymer degraded within 2 weeks, and clinical observations showed good biocompatibility of POE with no toxicity to the ocular tissues or increase in intraocular pressure. The injection of a larger volume, 100 microL, of POE, appeared inappropriate because of direct contact of polymeric material with the corneal endothelium, and triggered reversible edema and inflammation in the anterior chamber of the eye that regressed after a few days. After intravitreal administration, POE was well tolerated and no inflammatory reaction developed during the observation period. The polymer degraded slowly, appearing as a round whitish bubble in the vitreous cavity. The presence of modulators of degradation both improved POE biocompatibility and prolonged polymer lifetime in the eye. POE appears to be a promising biomaterial for clinical intraocular application.

A new mutation (A546T) of the betaig-h3 gene responsible for a French lattice corneal dystrophy type IIIA.

PURPOSE: To characterize the betaig-h3 gene defect in a French family affected with lattice corneal dystrophy type IIIA (LCDIIIA). METHODS: Histologic examination was performed from corneal buttons of two patients. Genomic DNA was extracted from leukocytes, and exons of the betaig-h3 gene were amplified by polymerase chain reaction to be directly sequenced. RESULTS: Numerous deposits were evident in the stroma and beneath the Bowman membrane, which had all the features of amyloid deposits. Analysis of exon 12 revealed a heterozygous G to A transition on codon 546. CONCLUSION: In contrast to Japanese patients, these French patients affected with LCDIIIA carry a distinct mutation of the betaig-h3 gene (A546T instead of P501T). Therefore, it is unclear whether different mutations could result in the same dystrophy or whether we are dealing with clinical heterogeneity of LCDIIIA.

Classical LCAT deficiency resulting from a novel homozygous dinucleotide deletion in exon 4 of the human lecithin: cholesterol acyltransferase gene causing a frameshift and stop codon at residue 144.

Lecithin: cholesterolacyltransferase (LCAT) transacylates the fatty acid at the sn-2 position of lecithin to the 3beta-OH group of cholesterol forming lysolecithin and the majority of cholesteryl ester found in plasma. LCAT participates in the reverse cholesterol transport pathway in man where it esterifies tissue-derived cholesterol following efflux from peripheral cells into HDL. Only 38 unique mutations in the human LCAT gene have been reported worldwide. Our French female proband presented with corneal opacity and no detectable plasma LCAT activity using either endogenous or exogenous assays. Her total plasma cholesterol and HDL cholesterol were low (2.34 mmol/l and 0.184 mmol/l, respectively) with a very high cholesterol/cholesteryl ester molar ratio (10.9:1). Plasma triglycerides were 0.470 mmol/l with low apo B (40.5 mg/dl), apo A-I (14.7 mg/dl), apo A-II (6.8 mg/dl) and apo E (2.1 mg/dl) levels. Plasma lipoprotein analysis by ultracentrifugation showed very low HDL concentrations and a characteristic shift of the lipoprotein profile towards larger, less dense particles. No proteinuria, renal dysfunction or signs of atherosclerosis were noted at age 45. Sequence analysis of her LCAT gene showed a novel homozygous TG-deletion at residues 138-139 that resulted in a frameshift causing the generation of a stop codon and premature termination of the LCAT protein at amino acid residue 144. Western blotting of the patient's plasma using a polyclonal IgY primary antibody against human LCAT failed to demonstrate the presence of a truncated LCAT protein. A 53 bp mismatched PCR primer was designed to generate an Fsp 1 restriction site in the wild type sequence of exon 4 where the mutation occurred. The 155 bp PCR product from the wild type allele produced a 103 bp and 52 bp fragment with Fsp 1 and no cleavage products with the mutant allele thus permitting rapid screening for this novel mutation.

Novel synthetic meshwork for glaucoma treatment. I. Design and preliminary in vitro and in vivo evaluation of various expanded poly(tetrafluoroethylene) materials.

A novel drainage implant for glaucoma filtering surgery (MESH) is proposed. After various expanded poly(tetrafluoroethylene) (e-PFTE) materials were evaluated, the feasibility and the short-term safety of the technique were assessed in this first pilot study in the rabbit. The porous structure and the in vitro resistance to aqueous flow of seven different e-PTFE membranes (5-80 microm average pore size) were compared. Eight Dutch pigmented rabbits were implanted with the T-shaped MESH implants made from either 20- or 50-microm average pore size e-PTFE membranes. Clinical examination, intraocular pressure (IOP) measurements, and histology analyses were performed over a period of 3 months. The contralateral nonoperated eyes served as controls. MESH implantation took less than 7 min. No postoperative hypotony, migration, or extrusion of the implant and no intraocular inflammation or infection occurred. A significant IOP reduction in the implanted eyes was obtained past postoperative day 21 with the 20-microm material implant. The drainage efficacy was correlated with the degree of colonization of the porous materials and the inner spacing of the implant as observed by histology. With a filtering patency 3 times longer than conventional trabeculectomy and laser sclerectomy, MESH surgery is a promising technique for glaucoma treatment. Further studies are underway to enhance the device efficacy and understand the mechanism of filtration.

Retinal TUNEL-positive cells and high glutamate levels in vitreous humor of mutant quail with a glaucoma-like disorder.

PURPOSE: To investigate whether retinal cell death observed in an avian glaucoma-like disorder occurs by apoptosis and whether an increase in excitotoxic amino acid concentration in the vitreous humor is associated temporally with cell death in the retina. METHODS: Presumptive retinal apoptotic nuclei were identified by histochemical detection of DNA fragmentation (by TdT-dUTP terminal nick-end labeling [TUNEL]), and vitreal concentrations of glutamate and several other amino acids were determined by high-pressure liquid chromatography with fluorometric detection in the al mutant quail (Coturnix coturnix japonica) in which a glaucoma-like disorder develops spontaneously. RESULTS: TUNEL-labeled nuclei were located mostly in the ganglion cell layer (GCL) in the retina of mutant quails 3 months after hatching. However, labeled nuclei were also observed in the inner and outer nuclear layers. At 7 months, most TUNEL-positive nuclei were detected in the inner nuclear layer, whereas labeled cells in the GCL were reduced in number. No TUNEL-labeled nuclei were detected in the retina of control quails at any age. Vitreal concentrations of glutamate and aspartate were significantly increased in 1-month-old mutant quails compared with control animals. Concentrations decreased at 3 months, and no significant differences were observed between strains at 7 months. CONCLUSIONS: Presumptive apoptotic cell death is detected from 3 months after hatching in mutant quails and is not restricted to retinal ganglion cells. Cell death appears just after a significant increase in excitotoxic amino acid concentrations in the vitreous humor, suggesting a correlation between both events.

[Artificial trabeculum (MESH). Clinical and histological study in the rabbit]. / Le trabéculum artificiel (MESH). Etude clinique et histologique chez le lapin.

PURPOSE: Designed to avoid postoperative hypotony that often occurs after trabeculectomy and to maintain long lasting filtration, the MESH is a thin porous expended polytetrafluoroethylene (ePTFE) implant that mimics the physiological meshwork. The aim of this study is to assess the tolerance, biocompatibility and effectiveness of this device during 6 months in the rabbit. MATERIAL AND METHODS: We used an ePTFE with about 5 microns pore size (Zytex). The head of the implant is 3,0 mm wide and 1,5 mm long and fits in the anterior chamber. The tail is 2,0 mm wide and 3,0 mm long and fits in the subconjunctival space. The MESH is 250 microns thick. 24 Dutch pigmented rabbits were selected because their dark pigmented iris contrasts with the ePTFE implant giving a better visualization. All the animals were cared for in accordance with ARVO resolutions. Surgery was performed on the right eye by the same surgeon (P.H.), the left eye serving as control for IOP measurements. The animals were distributed in 3 groups: one with MESH alone (MESH), one with MESH and Mitomycin C (MMC), one with MESH and 5-FU (5-FU). Follow-up was performed every week (W) during 6 months including IOP measurement, slit lamp observation, photography and bleb assessment. Histological study was done at POD 0, 15, 30, 90 and 180 one eye in each group. Student t test and alternate Welch t test were used for statistics. RESULTS: Filtering bleb: no bleb was visible before W3. A bleb was found between W3 and W6, decreasing between W6 and W9 with no more change after W10. The MESH implant: no change appears in the color of the MESH during the study. Some iris pigments or synechiae were seen in some cases. No extrusion occurred. Intraocular pressure: IOP was lower than in the control eye. The statistical analysis showed a significant lower pressure for the MESH alone at W5 (p = 0.0069), for the 5-FU group at W1 (p = 0.0326), W2 (p = 0.0488), W4 (p = 0.0312). With Mitomycine C we found very significant results at W1 (p = 0.0073), W2 (p = 0.0136), W4 (p = 0.0497), W9 and W11 (p = 0.0174). After W12 the groups were joined and IOP was significantly decreased at W17 (p = 0.0376) and W23 (p = 0.0462). Histology confirmed the correct position of the MESH, its biocompatibility and its ability to drain aqueous humor even if there is colonization of the pores by fibroblast-like cells. CONCLUSION: The present study has shown that the filtering bleb appeared after 3 weeks without major hypotony. The material was integrated only in the intrascleral tunnel and was stable. After 6 months the Mesh was well tolerated. The new concept has a simple surgical technique, less invasive than trabeculectomy and required less surgical time. This technique reduced IOP and produced long survival blebs in rabbits. This device appears suitable for the surgical treatment of open angle glaucoma.

Reduction of corneal edema in endotoxin-induced uveitis after application of L-NAME as nitric oxide synthase inhibitor in rats by iontophoresis.

PURPOSE: To investigate the involvement of the cornea during endotoxin-induced uveitis (EIU) in the rat and the effect of Ngamma-nitro-L-arginine methyl ester (L-NAME) as nitric oxide synthase (NOS) inhibitor, administered by iontophoresis. METHODS: EIU was induced in Lewis rats that were killed at 8 and 16 hours after lipopolysaccharide (LPS) injection. The severity of uveitis was evaluated clinically at 16 hours, and nitrite levels were evaluated in the aqueous humor at 8 hours. Corneal thickness was measured, 16 hours after LPS injection, on histologic sections using an image analyzer. Transmission electron microscopy (TEM) was used for fine analysis of the cornea. Transcorneoscleral iontophoresis of L-NAME (100 mM) was performed either at LPS injection or at 1 and 2 hours after LPS injection. RESULTS: At 16 hours after LPS injection, mean corneal thickness was 153.7+/-5.58 microm in the group of rats injected with LPS (n=8) compared with 126.89+/-11.11 microm in the saline-injected rats (n=8) (P < 0.01). TEM showed stromal edema and signs of damage in the endothelial and epithelial layers. In the group of rats treated by three successive iontophoreses of L-NAME (n=8), corneal thickness was 125.24+/-10.36 microm compared with 146.76+/-7.52 microm in the group of rats treated with iontophoresis of saline (n=8), (P=0.015). TEM observation showed a reduction of stromal edema and a normal endothelium. Nitrite levels in the aqueous humor were significantly reduced at 8 hours by L-NAME treatment (P=0.03). No effect on corneal edema was observed after a single iontophoresis of L-NAME at LPS injection (P=0.19). Iontophoresis of saline by itself induced no change in corneal thickness nor in TEM structure analysis compared with normal rats. CONCLUSIONS: Corneal edema is observed during EIU. This edema is significantly reduced by three successive iontophoreses of L-NAME, which partially inhibited the inflammation. A role of nitric oxide in the corneal endothelium functions may explain the antiedematous effect of L-NAME.

Confocal microscopy of cystic disorders of the corneal epithelium.

OBJECTIVE: This study aimed to describe the morphology of cystic disorders of the corneal epithelium by confocal microscopy. DESIGN: The study design was a prospective evaluation of confocal microscopic images of patients with cystic corneal disorders. PARTICIPANTS: Thirteen patients (19 eyes) were included. The corneal disorders included four patients with corneal decompensation (Fuchs' dystrophy), five patients with epithelial basement membrane dystrophy (e.g., Cogan's microcystic and map-dot dystrophies), one patient with Meesmann's dystrophy, and three patients with recurrent erosion syndrome of unknown etiology. Confocal images of diseased corneas were compared with those of ten normal control eyes (ten subjects). INTERVENTION: All patients were examined by slit-lamp biomicroscopic analysis and confocal microscopic analysis (Tomey, Erlangen-Temmenlohe, Germany). Image analysis was used to identify the corneal epithelial structures correlated with the corresponding pathology. MAIN OUTCOMES MEASURES: Confocal microscopy was used to assess the size, shape, light scatter, and reflection of the cysts. RESULTS: Slit-lamp examination results showed corneal epithelial cystic lesions in all cases. Confocal microscopy was able to identify cystic lesions in 9 (69.2%) of 13 patients. Of the four patients in whom lesions could not be found by confocal microscopy, three had recurrent erosion syndrome and the other one had epithelial basement membrane dystrophy. The confocal images were compatible with the clinical and histologic pictures of the disease. Normal control eyes did not show any epithelial lesion, either by biomicroscopy or confocal microscopy. CONCLUSIONS: Confocal microscopy provides an in vivo evaluation of cystic epithelial corneal lesions. This study shows that confocal microscopy is suitable for examining cystic lesions of the corneal epithelium. Nevertheless, it is not as sensitive as biomicroscopy in detecting cystic lesions in certain corneal conditions.

Endothelial damage caused by uncoated and fluorocarbon-coated poly(methyl methacrylate) intraocular lenses.

PURPOSE: To assess endothelial damage induced by poly(methyl methacrylate) (PMMA) intraocular lenses (IOLs) coated with a fluorocarbon polymer, Teflon AF, to make them highly hydrophobic. SETTING: Department of Ophthalmology. Hôtel-Dieu Hospital, Paris, France. METHODS: Ten Teflon-coated and 10 uncoated PMMA IOLs were used in an in vitro static touch model. The corneal endothelium was placed in direct contact with the IOL for 15 seconds and then stained with trypan blue and alizarin red. The endothelial damage produced by each IOL in the area of contact was assessed semiquantitatively and quantitatively. RESULTS: Teflon-coated IOLs produced significantly less endothelial damage than uncoated PMMA IOLs (P < .0001). Endothelial cells in contact with Teflon-coated IOLs did not usually adhere to the IOL surface. In contrast, the uncoated IOLs produced large areas of endothelial cell loss. CONCLUSION: Teflon-coated PMMA IOLs have an antiadhesive effect that reduced endothelial damage after IOL insertion in an in vitro model.

Influence of ePTFE polymer implant permeability on the rate and density of corneal extracellular matrix synthesis.

Microporous polymers have great potential for the production of corneal keratoprosthetic devices. Keratocytes invade the pores of expanded polytetrafluoroethylene implants (ePTFE) and collagen synthesis occurs. This ePTFE becomes translucent after its implantation in the stroma of rabbit cornea. The rate and density of cell growth within this polymer depends on the implant thickness, pore size, and its placement in the cornea. We have investigated the influence of the polymer permeability on the collagen and protein contents ePTFE implants. Rabbit corneal stroma were implanted with ePTFE disks (6 mm in diameter) by intralamellar keratoplasty. The implanted polymers were removed from the stroma after 3 to 6 months. The collagen and protein contents were determined after pepsin solubilization. The collagen content of the high-permeability implant was 3.7-fold greater than that of the low-permeability implant 3 months after implantation and 2.4-fold greater after 6 months. The total protein content of the high-permeability implant was 2.5-fold greater than that of low-permeability implant at 3 months and was the same after 6 months. The collagen-to-protein ratio was 68% in the high-permeability implants, and thus similar to that of normal corneal stroma. Thus, high polymer permeability increased both the rate and density of the corneal extracellular matrix ingrowth.

[Reticulated polyethylene oxide for gel injection adjustable keratoplasty. Biocompatibility in critical situation]. / Oxyde de polyéthylène réticulé pour kératoplastie ajustable par injection de gel (GIAK). Biocompatibilité en situation critique.

BACKGROUND: GIAK is a surgical technique developed for the correction of myopia. We evaluated the biocompatibility of a gamma-ray crosslinked poly(ethylene oxide) gel in two critical situations. MATERIALS AND METHODS: In 3 rabbits, a 5 mm inner diameter annular channel of 1 mm width was delaminated at 80% corneal depth and a sterile gamma-ray crosslinked polyethylene oxide gel was injected in the channel. In cornea 1 (control), the annulus was centered on the corneal apex and the channel was fully filled with gel. In cornea 2, the delamination was purposely decentered and the channel was approximately 1 mm from the limbus at its closest point. In cornea 3, the centered channel was purposely partially filled. After a 6 month follow-up, the rabbits were sacrificed for histology. RESULTS: Histopathology showed the oval channel walls to be irregular and a slight thinning of the epithelium located above the implant but no abnormalities in the stromal thickness. No mononuclear, polynuclear cells or other signs of encapsulation or rejection were found around the polymer in any of the animals. In cornea 3, fibroblasts were found on the lateral sides of the hydrogel globules. No limbal neovascularization was observed over the 6 month period in any of the animals. CONCLUSIONS: The injected PEO gel followed intimately the delaminated stromal channel and was well tolerated in the cornea. The fibroblastic reaction found in partially filled channels is caused by the normal wound healing process and not by the hydrogel chemical properties.