Jun Fu

A histopathologic study of retinal lesions inflicted by transscleral iontophoresis

In the present study, retinal lesions were induced by transscleral iontophoresis (1.5 mA) in rabbits. The size and severity of the lesions increased with the duration of application (2–25 min). No lesion was noted after <1 min application. Immediately after 5 min iontophoresis, the edematous retina exhibited necrotic retinal pigment epithelium (RPE), loss of outer segments, and thinning of the inner and outer nuclear layers. At 5 days after iontophoresis, there was a proliferation of RPE cells and macrophages in the subretinal space, with thinning of the inner and outer retinal layers continuing. By day 14, the retina had been reduced to a glial membrane. Immediately after 15 min iontophoresis, the damaged retina appeared in a mummified form containing no cellular elements. By day 5 thereafter, macrophages and actively proliferating RPE cells had been noted in the necrotic retina. By day 14, a glial membrane had formed.

A COMPARISON OF CONTINUOUS VERSUS INTERMITTENT LIGHT EXPOSURE ON APOPTOSIS

PURPOSE We recently found that continuous light exposure at a moderate intensity triggered apoptosis of photoreceptor cells. Since intermittent light exposure is known to cause more severe retinal damage than is continuous light exposure, we sought to determine if intermittent light exposure also triggered apoptosis of photoreceptor cells. METHODS Lewis albino rats were reared, for 2 weeks, in cyclic light and dark adapted for 24 hr before light exposure. Rats were exposed to intermittent light or continuous light for 6 or 9 hr, respectively. Light-exposed rats were killed by lethal injection at three timepoints: immediately after light exposure, after 6 hr of dark recovery following light exposure and after 24 hr of dark recovery following light exposure. Retinal damage after light exposure was evaluated by morphology, morphometry, the terminal transferase-mediated biotin dUTP nick end labeling (TUNEL) technique for identification of nicked/cleaved nuclear DNA and agarose gel electrophoresis of retinal DNA. RESULTS Evaluation of morphology confirmed that intermittent light exposure caused more photoreceptor cell damage than did continuous light exposure of the same duration and intensity. The TUNEL technique showed that photoreceptor nuclei contained nicked or cleaved DNA after either intermittent or continuous light exposure, although more TUNEL-positive nuclei were observed after intermittent exposure. Agarose gel electrophoresis of retinal DNA showed internucleosomal DNA fragmentation, which is associated with apoptosis in samples from intermittent light exposure. CONCLUSIONS These data demonstrated that intermittent light exposure triggered apoptosis in more photoreceptor cells than did continuous light exposure of the same intensity and duration.

Methylprednisolone therapy in laser injury of the retina.

The efficacy of methylprednisolone in argon-laser-induced retinal injury in primates was evaluated by clinical, histopathologic, and morphometric criteria. Methylprednisolone was given with a loading dose of 30 mg/kg followed by 5.4 mg/kg per hour in three different regimens: (1) starting 24 h before laser and continuing for 4 days; (2) starting immediately after laser and continuing for 4 days; and (3) starting immediately after laser and continuing for 8 h. Fundus photography, fluorescein angiography, and histologic examination showed significant beneficial effects of all three treatments compared to controls. Morphometrically, at the center of the lesion, the width of disrupted outer nuclear layer, the width of the affected RPE, and the percentage of residual photoreceptor nuclei confirmed the efficacies of treatment regimens 1 and 2, but not treatment regimen 3.

Dexamethasone ameliorates retinal photic injury in albino rats

The effect of dexamethasone in two regimens on retinal photic injury was studied in Lewis albino rats that were exposed to 24 hr of continuous green fluorescent light. Under regimen 1, dexamethasone was given at a daily dosage of 1 mg kg-1 for 8 days, starting 6 days before light exposure. Under regimen 2, dexamethasone was given at the same daily dosage for 3 days, started 1 day before light exposure. Pathologic study of the light-exposed retina, morphometric evaluation of the photoreceptor cell loss, cell counts of the macrophages in the subretinal space, and measurements of rhodopsin levels were undertaken in the dexamethasone-treated and control retinas at various times. The administration of dexamethasone in both regimens did not produce pathologic changes in the retina before light exposure, but rhodopsin levels were significantly lowered in both treated groups when compared to corresponding vehicle treated control animals. Under regimen 1, at 6 hr after light exposure, both the treated and the control groups showed comparable loss of photoreceptor cells, degeneration of the photoreceptor elements and retinal pigment epithelium, but a significantly lowered level of rhodopsin in the treated group was noted. At 6 days after exposure, the outer nuclear layer thickness, and the outer and inner segments showed significant preservation in the treated group. Also in the treated group, the number of macrophages was significantly reduced and the retinal pigment epithelial (RPE) vacuolation was markedly less. However, there was no difference in rhodopsin levels. At 14 days after exposure, the outer nuclear layer thickness and rhodopsin levels of the treated rats had significantly higher values than the controls. Under regimen 2, however, at 6 days after exposure, an ameliorative effect in the RPE was observed but there were no differences of rhodopsin levels, the outer nuclear thickness and number of macrophages between the treated and control groups. Regimen 1 was associated with a significantly higher retinal level of dexamethasone when compared with regimen 2. The ameliorative effect of dexamethasone on rat retinal photic injury may be through inhibition of lipid peroxidation, in which a high retinal level of the steroid is required.

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

Abstract• 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 methylprednisclones anti-inflammatory action, protection of the microcirculation and anti-lipid peroxidation effect. Proprietary interest: none

A Morphometric and Immunopathologic Study of Retinal Photic Injury in Primate

Clinical, histopathological, and ultrastructural studies examining the pathogenesis of retinal photic injury in animal models were performed by methods and yielded important qualitative information1,2. Morphometric studies of retinal photic injury in rodents had provided us with additional understanding of the more subtle pathophysiologic factors. For example, with morphometric techniques, it was determined that superior and temporal quadrants of the rodent retina arc more sensitive to retinal photic injury than the inferior and nasal quadrants3; aging rats are more susceptible to retinal photic injury4; light-adapted animals suffer less than the dark-adapted animals5; and different schedules of light exposure inflicted different severity of retinal lesions6. Hormonal influences on the severity of retinal lesions have also been defined7. Furthermore, with morpho-metric comparative methods, pharmacologic agents such as vitamin C8, dimethylthiourea9, flunarizine10, dexamethasone11, and methylprcdnisolone12 were demonstrated to rescue photoreceptor cells from light exposure.