Reattachment to a substrate prevents apoptosis of human retinal pigment epithelium

Background: Epithelial cells generally fail to survive in suspension. Harvesting human retinal pigment epithelium (RPE) for transplantation may separate the cells from their extracellular matrix and induce apoptosis. We investigated whether reattachment of RPE to a substrate will prevent apoptosis. Methods: Second-passage human RPE cells were plated onto tissue culture plastic precoated with extracellular matrix, fibronectin or laminin, uncoated tissue culture plastic, untreated plastic and untreated plastic coated with 4% agarose. Reattachment rates were determined for each substrate 24 h after plating. The TUNEL technique was used to determine apoptosis rates in attached cells, unattached cells and the entire cell population. Results: Attachment rates were as follows: ECM-coated tissue culture plastic fibronectin-coated tissue culture plastic laminin-coated tissue culture plastic uncoated tissue culture plastic untreated plastic agarose-coated untreated plastic. Apoptosis rates for the entire cell population increased as the RPE cell attachment rate decreased. The proportion of apoptotic cells in the entire population was inversely related to the percent attached cells (r = -0.95). Conclusion: Reattachment of harvested RPE to a substrate decreased the rate of RPE apoptosisin vitro. RPE cells which are removed from their substrate prior to transplantation must reattach rapidly to a substrate to prevent apoptosis.Presented in part at the Annual Meeting of the Association for Research in Vision and Ophthalmology, Fort Lauderdale, Florida, May 1996     

Unusual retinal pigment epitheliopathy and choroidopathy in carcinomatosis: A rare case of cancer-associated retinopathy

Background: Cancer-associated retinopathy is a syndrome causing ocular symptoms. It is a rare entity and only a few cases have been reported. Methods: A 67-year-old woman with small-cell endometrial carcinoma suffering from deterioration of visual acuity is presented. Results: The patient presented with extensive mottled changes of the retinal pigment epithelium, accompanied by diffuse subretinal fluid in the posterior pole and exudative retinal detachments inferior in both eyes. Conclusion: This patient suffered from a rare variety of cancer-associated retinopathy.     

Isovolemic hemodilution increases retinal tissue oxygen tension

Background: Therapeutic isovolemic hemodilution has been reported to improve blood flow and oxygen delivery. Few reliable measurements have been made, however, showing the effect of hemodilution on tissue oxygen tension. Methods: We measured retinal oxygen tension during experimental isovolemic hemodilution in normal cats. Polarographic oxygen microelectrodes were placed in the vitreous humor within 100–200 m of the retinal surface.Results: Tissue oxygen tension increased initially during isovolemic hemodilution to a maximum approximately 50% above baseline at approximately two thirds of the original hematocrit level. Hemodilution beyond this point to lower hematocrits caused a steady decline in tissue oxygen tension. Cardiac output measured in one cat undergoing isovolemic hemodilution increased as hematocrit was lowered, but the cardiac erythrocyte flux actually decreased steadily.Conclusion: The observed increase in tissue oxygen tension with hemodilution appears to be explained by a lesser reduction in capillary than in systemic hematocrit, coupled with an increased capillary blood flow. The increase in tissue oxygen tension we observed could in part explain the clinically beneficial effects of hemodilution.     

Effects of norfloxacin on the retina in rabbits

Background: Fluoroquinolones have a strong affinity with melanin, and their ocular effects have been reevaluated. Norfloxacin, one of the fluoroquinolones, has broad-spectrum activity against aerobic gram-positive and gram-negative bacteria. We examined the retinal toxicity and intraocular pharmacokinetics of intravitreal norfloxacin in rabbits. Methods: Twenty-three albino and 23 pigmented rabbits were divided into three groups to evaluate retinal toxicity and two groups to investigate the intraocular pharmacokinetics. Each of these five groups was further divided into two subgroups (albino rabbits and pigmented rabbits). Results: With 500 Etg norfloxacin, the oscillatory potential of the electroretinogram was transiently and selectively deteriorated in albino and pigmented rabbits, whereas the electroretinogram remained unchanged with 50 g in pigmented rabbits. No changes were observed in the visual evoked potential or on histology of the retina 7 days after an intravitreal injection of 50 or 500 ltg norfloxacin. The electroretinogram and the retinal histology became abnormal 7 days after four intravitreal injections of 500 g norfloxacin at 7-day intervals. As regards the intraocular pharmacokinetics after an intravitreal injection, the norfloxacin concentration in the chorioretina was as high as that in the vitreous 3 h after injection and was much higher than that in the vitreous 7 days after injection. Similar results were obtained after multiple injections. Conclusion: These results indicate a high concentration of norfloxacin in the melanin-containing ocular tissues.     

The effect of nasolacrimal occlusion on drug-induced mydriasis

Background: Nasolacrimal occlusion has been shown to improve the efficacy of some topically applied ocular drugs. The aim of this study was to investigate the effect of nasolacrimal occlusion on tropicamide-induced mydriasis. Methods: We compared pupillary dilatation by 0.125% tropicamide with and without nasolacrimal occlusion in 40 healthy volunteers. Results: Analysis of variance with repeated measures failed to show any advantage due to nasolacrimal occlusion in drug-induced mydriasis. Conclusion: Nasolacrimal occlusion did not increase the mydriasis obtained with 0.125% tropicamide.     

The retinal depression sign in diabetic retinopathy

Background: The retinal depression sign, an abnormal light reflex due to ischemia-induced focal inner retinal atrophy, was initially described in patients with sickle cell retinopathy. We undertook this study to characterize the finding in diabetic retinopathy. Methods: The fundus photographs of 97 consecutive patients with diabetic retinopathy seen in the Penn State University Ophthalmology Department were reviewed. We recorded age, sex, type of retinopathy, visual acuity and photographic details, including the presence or absence of the retinal depression halo. Results: The retinal depression sign was present in 9 of 22 patients age 45 years or under, versus 1 of 75 patients age 46 or older (P<0.0001, two-tailed Fisher's exact test). The presence or absence of retinal depression sign did not predict the type of retinopathy in a given patient. Eleven (70%) of the affected eyes had visual acuity 20/40. Conclusion: The retinal depression sign is common in younger patients with diabetes and should be distinguished from other macular lesions associated with diabetic retinopathy.     

Indocyanine green and fluorescein angiography of surgically excised macular choroidal neovascularizations: correlations with histopathologic and ultrastructural findings

Background: Informations are expected from the histopathological study of surgically excised choroidal neovascular membranes (CNMs), particularly in relation to the angiographic aspects of vascular architecture. Methods: Fluorescein and indocyanine green (ICG) angiograms were studied together with the histopathological features of 12 surgically excised subfoveal CNMs in eyes affected by age-related macular degeneration (ARMD). Results: Instead of the early and delayed diffuse hyperfluorescence secondary to CNMs observed on fluorescein angiography (seven were well defined, five scar evolved), ICG revealed (a) very early hypofluorescence of the membrane bulk over the fluorescence of the outer choroidal vascular bed and (b) late hyperfluorescence gradually increasing and partially defining the border of membranes. CNMs with well-defined hyperfluorescent aspects were characterized by fibrovascular bulk lined on one side by retinal pigment epithelium. Fibrosis reaction predominated over the vascular components in scar-evolved membranes. Conclusions: Fluorescein and ICG angiographic differences in the appearance of CNMs could depend on (a) the morphological structure and size of the CNM (b) its location within the chorioretinal layers and (c) different molecular characteristics of the dyes used.     

Ocular blood flow velocity reduction after buckling surgery

Background: It has been reported that scleral buckling reduces the blood flow velocity in retinal vessels. Blood flow changes may also appear in other ocular and extraocular vessels. This study describes the blood flow velocity changes in the ophthalmic artery (OA) after performing this procedure. Methods: The study was carried out in 12 patients (12 eyes) with rhegmatogenous retinal detachment. Color Doppler imaging was used to measure the peak and average blood flow velocity in the OA. Measurements were taken 1 day before and 2 days after scleral buckling surgery was performed. Intraocular pressure (10P) was measured prior to each ultrasound study. Results: We found that statistically significant reductions in the peak flow velocity (33%) and average flow velocity (31%) occur in the OA after scleral buckling. All patients showed an increase in IOP after surgery. Conclusion: Buckling surgery reduces the blood flow velocity in the OA. Since the OA is the origin of the arterial branches that supply blood to the eye, our results suggest that scleral buckling may decrease not only retinal but also choroidal blood perfusion. Some extraocular structures might also be affected.     

Ophthalmological and neuro-ophthalmological involvement in Churg-Strauss syndrome: a case report

Background: It is well known that different types of eye involvement may develop during the course of systemic vasculitides. Methods: We report here a case of Churg-Strauss syndrome (allergic granulomatous angiitis) characterized by the presence of multiple ophthalmological and neuro-ophthalmological lesions, i.e., mononeuritis of the fourth cranial nerve, multifocal choroidal ischaemia, and bilateral ischaemic optic neuropathy. Results: Ischaemic lesions in the posterior ciliary plexus and chorio-retinal circulation, which appeared simultaneously after a phase of disease activity, were documented. Conclusion: The simultaneous occurrence of multiple ocular features in a patient with Churg-Strauss syndrome suggests that regional vasculitis may be the pathological mechanism underlying the multiple ophthalmological lesions in this disorder.     

Anterior chamber inflammation after transconjunctival cryosurgery

Background: Inflammation caused by transconjunctival cryotherapy for prophylactic retinal detachment surgery was measured in various conditions. Methods: Thirty-four eyes of 28 patients with peripheral retinal lesions predisposing to retinal detachment were studied by laser flare cell meter before and after treatment. Results: The mean flare value for 34 eyes was 4.06 ± 1.45 photon counts/ms before surgery and 5.72 ± 2.52 pc/ms after surgery (p < 0.05). Flare value was elevated at 1, 2, and 3 weeks after treatment, peaking at 2 weeks (p < 0.05), and normal again at 4 weeks. There were no significant differences in flare increase between eyes with and without retinal breaks, eyes with and without limited retinal detachment, eyes with myopia more and less than – 8.0 D, and eyes with a treatment area limited to one quadrant and extending over more than one quadrant. Conclusion: Transconjunctival cryosurgery caused mild inflammation in the anterior chamber of the eye for 3 weeks. The inflammation was not affected by the presence of retinal break or limited retinal detachment, the degree of myopia, or the extent of the treatment area.     

The effect of high-dose methylprednisolone on laser-induced retinal injury in primates: an electron microscopic study

Background: Previously we reported an ameliorative effect of high-dose methylprednisolone in laser injury to monkey retinas. The ultrastructural modification by methylprednisolone has not been examined. Methods: Cynomolgus monkeys were given severe kgrade III) retinal laser burns and treated with an intravenous megadose of methylprednisolone. Pathologic features of the retinal lesions with or without methylprednisolone treatment were evaluated by light and electron microscopy. Results: Ultrastructurally, the treated lesions showed rapid recanalization of choriocapillaris; proliferation of retinal pigment epithelium to replace the necrotic and damaged cells, resulting in rapid re-establishment of blood retinal barrier; mild macrophagic activity; and rapid reformation of the outer limiting membrane by Mueller cells. Conclusion: A high dose of methylprednisolone affected the responses of the choriocapillaris, retinal pigment epithelium, photoreceptor cells and Mueller cells to laser injury, showing an overall beneficial effect. These modifications might be ascribed to methylprednisclone's anti-inflammatory action, protection of the microcirculation and anti-lipid peroxidation effect. Proprietary interest: none     

Morphometry of the optic nerve and retinal vessels in children by computer-assisted image analysis of fundus photographs

Background: The retinal fundus in childhood has a different morphology than in adulthood. Existing methods are not suitable for evaluation of fundus photographs from children. Therefore, a new method for quantitative analysis of fundus morphology utilizing a personal computer-assisted digital mapping system was developed. Methods: A CCD flatbed scanner is used to digitize fundus photographs, producing computer images which are analyzed on an IBM/AT computer. Area measurements of the optic disc, excavation and peripapillary crescent are made, as well as determinations of the length, branching, tortuosity and distribution of the retinal vessels on the fundus surface. Results: Determination of the inter-and intra-observer variability of the computer-assisted image analysis technique demonstrated good reproducibility. The method is demonstrated using fundus photographs of six normal children and six children with the fetal alcohol syndrome. Typical variations in appearance of optic disc and retinal vessels are seen. Conclusion: The system is unique in measuring both the optic nerve head and the retinal vessels and is therefore especially useful for detailed studies of normal and abnormal development of these structures in children.     

Choroidal metastasis of mixed carcinoma of the parotid gland

Background: A case of mixed carcinoma of the parotid gland (an epidermoid carcinoma located in a pleomorphic adenoma) metastatic to the choroid is presented. The histopathology of the tumor is discussed. Methods: A 65-year-old man was admitted complaining of blurred vision in his right eye for 1 day. He underwent parotidectomy for mixed carcinoma of the parotid gland (an epidermoid carcinoma located in a pleomorphic adenoma that was completely excised) 6 months before. Funduscopic examination showed a nasal retinal detachment, with gray-whitish, minimally elevated nodular choroidal lesions. Fluorescein angiography and contact B-scan ultrasonography confirmed the presence of an underlying mass. Results: The right eye was enucleated and an epidermoid infiltrating carcinoma was identified. Conclusions: Metastatic tumors are the most common intraocular malignancies, and the choroid is by far the most common location for intraocular metastases. There are few cases reported of parotid tumors metastatic to the orbit. To the best of our knowledge, no histological examination of an ocular metastatic mixed carcinoma of the parotid gland has yet been reported.     

Blood-retina barrier in acute retinal branch vein occlusion

Background: Extensive clinical studies on retinal branch vein occlusion have not yet been able to clarify its pathogenesis. A study designed to look at the associated blood-retina barrier changes may contribute to a better understanding of the different forms of evolution of this pathology. Methods: A prospective study was done in seven patients with recent large temporal branch vein occlusion. Vitreous fluorophotometry, fluorescein angiography and retinal colour photography were performed within the 1st week after the onset of symptoms, 1 week later, and at 12 and 24 weeks. Results: A more marked blood-retina barrier breakdown was found at 1, 2, 12 and 24 weeks in the eyes that later developed extensive capillary non-perfusion. Conclusions: Our results suggest that the breakdown of the blood-retina barrier may play an important role in the subsequent development of retinal nonperfusion in eyes with large branch vein occlusion. We postulate that the eyes that will present later extensive capillary nonperfusion develop, from the initial stages of the disease, a progressive ischaemic capillaropathy characterized by blood-retina barrier breakdown. Retinal pigment epithelium degeneration and arterial lumen narrowing, secondary to the vein obstruction, may help to increase and perpetuate the blood-retina barrier breakdown during the first 6 months after the occlusion.     

Indocyanine green videoangiographic findings in choroidal metastatic tumor

Background: Indocyanine green videoangiography (ICGV) has recently advanced and become widely available as a clinical tool. We tested the efficacy of ICGV in evaluating metastatic choroidal tumors. Methods: ICGV using Topcon 50-IA was performed in five patients with clinically diagnosed choroidal metastatic tumors. The findings were compared with those of conventional fluorescein angiography (FAG). Results: Compared with FAG, ICGV demonstrated more smooth and regular hypofluorescent lesions, precisely indicating the exact size of the tumor. In all cases, no tumor vessels were found. The choroidal vascular integrity around the tumors was observed. FAG, however, was more sensitive in detecting the tumor development than ICGV when the extent of the hypofluorescent lesion could be seen clearly in the early phase. FAG provided more information regarding retinal pigment epithelial dysfunction in the outer blood retinal barrier overlying the tumor. Conclusion: Since ICGV enables visualization of tumors through the retina, it is very useful, particularly in cases of associated secondary retinal detachment around the tumors. ICGV combined with FAG provides more precise assessment of the tumors themselves and their response to treatment than FAG alone.     

Correlation of histological tumor vascularization and Doppler sonography in patients with malignant melanoma of the choroid

Background: In previous studies Doppler sonography was proven to the useful for diagnostics and follow-up of malignant melanomas of the choroid.Methods: To evaluate the correlation between Doppler sonographical findings and the histological tumor vascularization, we examined five eyes of five patients with malignant melanoma of the choroid before enucleation with an ATL Ultramark-8 Duplex scanner and compared these results with computerized planimetry of histological tumor vascularization after immunohistochemical staining of vascular endothelium with anti-factor-VIII antibodies.Results: We found no definite correlation of flow velocities with histological parameters of tumor vascularization. The tendency was for a decrease in maximum flow velocities as histological vascularization increased.Conclusions: Due to (1) the usual lack of sensitivity parameters of the used Duplex device for the clinical user and (2) the unpredictable branching pattern within a melanoma resulting in an unknown angle of incidence of the Doppler beam, we conclude that the quantitative results of Doppler sonography require cautious interpretation. Nevertheless, presence or absence of Doppler shifts is a valuable parameter for the follow-up of irradiated tumors.Poster presented at the annual meeting of the Association for Research in Vision and Ophthalmology, Sarasota, Florida, 1992     

Primary vitrectomy without scleral buckling for rhegmatogenous retinal detachment

Background: Pars planta vitrectomy has evolved as an alternative method in the treatment of more complicated rhegmatogenous retinal detachments. We report a series of patients who underwent primary vitrectomy with gas tamponade without the use of additional scleral buckling. Methods: A retrospective study of 53 patients with a follow-up of 6–45 months (mean 17.8 months) was carried out. Preoperative findings included unusual, multiple or large breaks, vitreous haemorrhage, proliferative vitreoretinopathy and bullous retinal detachment. Preoperative visual acuity was between light perception and 1.0, with 30% (16/53) of patients with 0.4 or better. Results: Retinal reattachment was achieved in 64% of cases (34/53) with one and in 92% (49/53) with one or more operations. Final visual acuity was between light perception and 1.0, with 41% (22/53) of patients with 0.4 or better. Cataract formation occurred in 86% (37/43) of all patients with a clear lens preoperatively. Macular pucker was noted in 11 % (6/53) and postoperative proliferative vitreoretinopathy causing redetachment in 6% (3/53). Conclusion: With primary vitrectomy, a high final anatomical success rate with few intraoperative complications can be achieved in more complicated forms of rhegmatogenous retinal detachment. The major drawback of the procedure is the high incidence of post-operative cataract formation.     

Tolerance of human fetal retinal pigment epithelium xenografts in monkey retina

Background: RPE transplantation offers the possibility of treating certain forms of retinal degeneration. Understanding how to optimize the surgical technique for performing RPE transplantation, especially in primates, is therefore of considerable interest. Methods: Fifteen patch RPE transplants were performed in six monkeys. The transplant sites were examined at follow-up by ophthalmoscopy, biomicroscopy, fluorescein angiography and histology. Foveal and peripheral retinal transplants were compared. Results: Human fetal RPE xenografts can survive without rejection for at least 6 months after transplantation in monkey retina. Such grafts form a basal lamina and make intimate contacts with the outer segments of the host. Both rods and cones retain a normal appearance when in contact with unrejected transplants. Rejection occurred in only 30% (3/10) of the peripheral but in 60% (3/5) of the foveal transplants. Conclusions: Cultured human fetal RPE patch transplants can survive and maintain local photoreceptor integrity for relatively long periods of time in monkey subretinal space without immunosuppression. Rejection, when it occurs, is more frequent near the fovea.     

Color Doppler imaging of the ocular vessels

Background: Color Doppler imaging allows for simultaneous two-dimensional anatomical imaging and Doppler evaluation of blood flow velocity. Methods: We examined 40 normal eyes (17 males and 23 females, aged 16 to 57 years) with this technique. The color Doppler unit used in this study had a 5.0-MHz PLF-503 ST phased-array scanning head. Each vessel examination was repeated 10 times during a single session. Results: The following peak flow velocities were found: central retinal artery, 12.5±2.4 cm/s; central retinal vein, 4.4±0.49 cm/s; posterior ciliary arteries, 14.4±2.6 cm/s; vorticose veins, 5.5±1.0 cm/s; ophthamic artery, 36.9±7.0 cm/s. Ophthalmic artery systolic and end-diastolic velocities declined as a function of age; however, these changes were not significant (systolic: r=–0.24; diastolic, r=–0.22). Conclusion: This noninvasive technique allows quantitative assessment of blood flow velocity in these vessels.     

Pressure sore in a patient who underwent repair of a retinal tear with gas injection

Purpose: To demonstrate a pressure sore following strict head positioning in a patient who underwent encircling band, vitrectomy and gas injection. Methods: A male patient was admitted to the hospital with a large posterior horseshoe tear in the inferior temporal retina with severe vitreous traction and retinal detachment. Encircling band, vitrectomy cryotherapy and gas injection was performed. After surgery the patient was instructed to sit in a facedown position. Results: A pressure sore resulted from prolonged immobility of the right elbow due to face-down positioning following encircling band, vitrectomy and gas injection. Conclusion: A patient injection. Conclusion: A patient may rarely have compulsive personality traits that result in extreme compliance to the physician's recommendations; therefore, general instructions given for head positioning should include permission for a change in position when required, at least for brief periods of time.