bromochloroacetic-acid and Retinal-Perforations

To elucidate the pathogenesis of macular hole formation, focusing in particular on the possible role of cellular migration on the cortical vitreous and internal limiting membrane (ILM) around the macular hole.. To gain a comprehensive overview of the ILM excised in macular hole surgery (n = 36), the ILMs were carefully unfolded and spread out onto glass slides as continuous flat sheets that each contained a macular hole. The specimens were observed by light microscopy and transmission electron microscopy (n = 9), and the cellular distribution was analyzed by scanning electron microscopy in a quantitative manner (n = 27). Immunohistochemistry for glial fibrillary acidic protein and cytokeratin 18 was carried out for cellular characterization. Cellular proliferation was assessed by immunohistochemistry for proliferating cell nuclear antigen and Ki-67.. Cellular migration was not apparent around the macular hole in the early stage of development of the macular hole (stage 2, 0 microm). As the macular hole passed through the later stages of development, cellular migration developed around the macular hole (stage 3, 84 microm) and the area of cellular migration gradually enlarged (stage 4, 420 microm). The immunophenotypic analysis showed that these cells were mainly glial fibrillary acidic protein-positive glial cells and cytokeratin 18-positive retinal pigment epithelial cells. The proliferating cell nuclear antigen and Ki-67 immunohistochemistry showed that some of these cells were proliferating on the ILM.. Cellular migration on the ILM is not necessary for the initial formation of a macular break. Cellular migration developed after the macular break occurred, and the migration and proliferation increased gradually from the macular hole.. This study provides a new method for understanding the ultrastructural analysis of the pathogenesis of the macular hole.

Topics: Basement Membrane; Cell Movement; Cell Proliferation; Epiretinal Membrane; Glial Fibrillary Acidic Protein; Humans; Keratins; Ki-67 Antigen; Microscopy, Electron, Scanning; Microscopy, Immunoelectron; Proliferating Cell Nuclear Antigen; Prospective Studies; Retinal Perforations

To understand molecular events that lead to retinal pigment epithelial (RPE) cell proliferation and migration during the early phases of proliferative vitreoretinopathy (PVR) in a rabbit model.. Retinal holes were created and interleukin-1beta(IL-1beta) was injected intravitreally. Eyes were examined by indirect ophthalmoscopy and eyecup pieces containing retinal holes were analyzed at different times after the surgery up to 4 weeks. RPE proliferation and migration were examined by immunohistochemistry. Tyrosine phosphorylation of extracellular signal regulated kinase (ERK) and hepatocyte growth factor receptor (HGFR or c-met) was determined by immunoprecipitation and western blot analysis. Tyrosine phosphorylation of c-met and morphological studies was performed on vitreous treated ARPE-19 cells. Expression of c-jun was determined by Northern blot analysis. Matrix metalloproteinase (MMP) content in vitreous was assessed by zymography.. Indirect ophthalmoscopy identified formation of epiretinal membrane and immunohistochemistry identified proliferative and migratory RPE and other cells in the posterior segment containing retinal holes at 4 weeks post-surgery. Tyrosine phosphorylation of ERK and c-met occurred in this segment within 30 min of surgery. ARPE-19 cells treated with vitreous from the 24 h post-surgical eyes, but not with control vitreous or IL-1beta, showed morphological changes and tyrosine phosphorylation of c-met. Northern blot analysis in this segment identified upregulation of c-jun within 30 min of surgery and the expression peaked at 72 h. Zymographic analysis of vitreous identified MMP-9 in 12-72 h post-surgery.. These data suggest that the presence of retinal holes and IL-1beta may lead to activation of HGF, mitogen activated protein kinases (MAPK), c-jun and extracellular matrix remodeling, resulting in proliferative and migratory cells in the wounded retina.

Topics: Animals; Blotting, Northern; Blotting, Western; Cell Division; Cell Movement; Disease Models, Animal; Hepatocyte Growth Factor; Interleukin-1; Keratins; Matrix Metalloproteinases; Mitogen-Activated Protein Kinases; Phosphorylation; Pigment Epithelium of Eye; Precipitin Tests; Proliferating Cell Nuclear Antigen; Proto-Oncogene Proteins c-jun; Proto-Oncogene Proteins c-met; Rabbits; Retinal Perforations; Tyrosine; Vitreoretinopathy, Proliferative; Vitreous Body

To immunocytochemically characterize the neural and glial elements of idiopathic full-thickness macular hole (FTMH) opercula excised during vitrectomy, and to correlate them with the outcome of surgery.. Opercula were collected from eyes undergoing vitrectomy for stage 3 FTMH and processed for transmission electron microscopy, light epifluorescence, and laser scanning confocal microscopy. Glia were identified using anti-glial fibrillary acid protein (GFAP), antivimentin, and anti-cellular retinaldehyde binding protein antibodies. Anti-phosphodiesterase gamma and antirhodopsin were used for cone and rod photoreceptors, and anticytokeratin was used for retinal pigment epithelium. The findings were correlated with the clinical data before and after surgery. For statistical analysis, data were combined with those of a previous study by the authors of 18 opercula.. Opercula from 12 consecutive eyes of 12 patients were studied. In all opercula, GFAP, vimentin, and cellular retinaldehyde binding protein-positive glia were present. Six (50%) of 12 opercula contained more than 5 photoreceptors with somata and internal photoreceptor fibres, but lacking outer segments, demonstrating strong immunoreactivity to anti-phosphodiesterase gamma without antirhodopsin reactivity consistent with cones. Further, 2 (17%) of 12 opercula showed few cones (1-5 cones), and 4 (33%) of 12 contained only glia. Clinicopathologic correlation of the 30 opercula from the 2 studies showed that eyes with opercula containing more than 5 photoreceptors were associated with a worse anatomical closure rate after initial surgery, compared with those with fewer than 5 photoreceptors (P =.004). Once closure had been achieved with reoperation, median postoperative vision was similar in both groups (20/40 and 20/60, respectively).. A spectrum of opercula occur in FTMH ranging from those containing only glia to those containing numerous cones. The extent of foveal neuroretinal tissue loss may affect the outcome of surgery.

Topics: 3',5'-Cyclic-GMP Phosphodiesterases; Aged; Carrier Proteins; Cyclic Nucleotide Phosphodiesterases, Type 6; Female; Fluorescent Antibody Technique, Indirect; Glial Fibrillary Acidic Protein; Humans; Keratins; Male; Microscopy, Confocal; Microscopy, Fluorescence; Middle Aged; Neuroglia; Photoreceptor Cells, Vertebrate; Retinal Perforations; Rhodopsin; Vimentin; Vitrectomy

To report an unusual case of intravitreal inflammation in a human eye caused by the presence of residual perfluorodecalin in a case of giant retinal tear and retinal detachment.. The posterior capsule of the lens, which was infiltrated with deposits, was collected during surgery. The specimen was stained with hematoxylin and eosin, with periodic acid-Schiff, and for melanin. Part of it was examined with electron microscopy. Immunohistochemical staining was performed to demonstrate CD68 antigens, cytokeratin, and glial fibrillary acid protein.. Vacuolated macrophages and retinal pigment epithelial cells infiltrated the posterior capsule. Electron microscopy showed the presence of membrane-lined vacuoles within the macrophages. A monolayer of epithelial cells covered the cellular infiltration.. Residual perfluorodecalin can induce an intraocular chronic macrophage response.

Topics: Adult; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Cryosurgery; Endophthalmitis; Eye Diseases; Fluorocarbons; Glial Fibrillary Acidic Protein; Humans; Immunoenzyme Techniques; Keratins; Lens Capsule, Crystalline; Macrophages; Male; Phacoemulsification; Reoperation; Retinal Detachment; Retinal Perforations; Scleral Buckling; Vitrectomy; Vitreous Body

To investigate the healing process of retinal holes, including the identification of the cell types which play an important role in the process, we created experimental retinal holes with minimal damage to retinal pigment epithelium (RPE) in rabbit eyes.. Pars plana vitrectomy was performed in the rabbit eye. A dome-shaped retinal detachment (bleb; diameter 1.5 mm) was made by injecting balanced salt solution into the subretinal space, followed by making a retinal hole (diameter 0.5 mm) in the center of the bleb with a silicone-tipped extrusion needle. In one group of rabbits, fluid-air exchange was performed and sulfur hexafluoride gas was injected into the vitreous cavity postoperatively. In another group, gas tamponade was not performed. The operated eyes were examined ophthalmoscopically and enucleated at 1, 4, 7, 14, 30, and 90 days after surgery. Tissues were prepared in 5-microm sections for hematoxylin-eosin staining and immunohistochemistry with antibodies to cytokeratin 18 and glial fibrillary acidic protein (GFAP) and examined by light microscopy.. In the gas-injected eyes, the retinal holes were ophthalmoscopically closed by 7 days after the surgery. Microscopic examination revealed that the sensory retina around the retinal hole was reattached, and the area of retinal defect was covered with cells which were positive for cytokeratin 18 and GFAP by 7 days after the surgery. In the eyes without gas tamponade, the retinal holes did not close during the observation period.. These findings suggest that early attachment between the sensory retina and RPE could be essential for closure of a retinal hole, where glial and RPE cells might play an important role. This model seems to be useful to investigate the process of closure of retinal holes.

Topics: Animals; Biomarkers; Cell Division; Disease Models, Animal; Glial Fibrillary Acidic Protein; Immunoenzyme Techniques; Keratins; Pigment Epithelium of Eye; Rabbits; Retina; Retinal Perforations; Sulfur Hexafluoride; Wound Healing

The standard treatment for retinal breaks is thermal adhesion. Breaks in the posterior pole (i.e., macular holes) recently have been treated using vitrectomy and the recombinant cytokine transforming growth factor-beta. This has been shown to achieve closure of the retinal breaks by stimulating localized fibrocellular proliferation. Serum has been shown to contain chemoattractants and mitogens for many types of cells. The authors studied the clinical and histologic effect of autologous serum application to retinal breaks in an experimental model.. Twenty-four rabbits underwent pars plana lensectomy, vitrectomy, retinectomy, fluid-air exchange, application of test solution (12 with Hank's buffered salt solution and 12 with autologous serum), and air-gas exchange. Clinical examination with indirect ophthalmoscopy was performed, and animals were killed 5, 14, and 28 days after treatment. Tissue sections through the retinectomy were studied by light microscopy, electron microscopy, and immunocytochemistry.. None of the serum-treated eyes showed retinal detachment at the site of the retinectomy by evaluation with indirect ophthalmoscopy at each of the time points. In contrast, in control eyes retinal detachment developed at the retinectomy site from 0% at day 5 to 50% at day 14 and 75% at day 28. By light microscopy, serum-treated eyes contained multilayers of fibroblast-like cells adhering the retinectomy edges to the underlying retinal pigment epithelium and choroid. The control eyes had nonadherent retinal edges at the retinectomy site with little sign of fibrocellular response. Results were confirmed by electron microscopy. The fibroblast-like cells by immunocytochemistry contained vimentin, cytokeratin 18, and/or glial fibrillary acidic protein.. This study suggests that serum induces a localized fibrocellular response at the retinectomy edges compared with control eyes. This response, characterized by light microscopy, electron microscopy, and immunocytochemistry, appears to involve a mixed population of glial, retinal pigment epithelial, and/or fibroblastic cells. These cells seem to enhance adhesion and subsequent reattachment of the edges of the retinectomies at the time points studied when compared with controls.

Topics: Animals; Blood; Cell Movement; Disease Models, Animal; Fibroblasts; Glial Fibrillary Acidic Protein; Immunoenzyme Techniques; Keratins; Male; Neuroglia; Pigment Epithelium of Eye; Rabbits; Retina; Retinal Detachment; Retinal Perforations; Vimentin

We performed electron immunocytochemical staining for cytokeratins and glial fibrillary acidic protein on cortical vitreous obtained at the time of vitrectomy from two patients with premacular hole lesions. The specimens were thought to represent cortical vitreous because each consisted of a sheet of tissue that, in addition to being firmly attached around the foveal lesion, was attached around the disc and extended well into the periphery. The specimens contained a moderate number of cells and an abundant extracellular matrix. Most of the cells were found singly or in small clusters on the surface of the matrix. Preembedding immunostaining on one specimen showed several cells that stained for cytokeratins and several that stained for glial fibrillary acidic protein. The majority of the cells on the matrix appeared to express one of these two intermediate-filament proteins. Postembedding immunogold double labeling on both specimens showed that most cells were labeled for either glial fibrillary acidic protein or cytokeratins, but there were a few cells that unequivocally expressed both proteins simultaneously. These data suggest that the cortical vitreous of patients with some premacular hole lesions contains retinal pigment epithelial and glial cells that may contribute to abnormal vitreoretinal adherence and could play a role in macular hole formation.

Topics: Aged; Female; Fluorescein Angiography; Fundus Oculi; Glial Fibrillary Acidic Protein; Humans; Immunoenzyme Techniques; Keratins; Microscopy, Immunoelectron; Retinal Perforations; Vitrectomy; Vitreous Body