Study of inhibitory effects of an antiangiogenic somatostatin-camptothecin conjugate on laser-induced choroidal neovascularization in rats.

PURPOSE: To evaluate the ocular toxicity and efficacy of intravitreal camptothecin-somatostatin conjugate (JF-10-81), a somatostatin type 2 receptor-directed, antiangiogenic compound. METHODS: Part 1: New Zealand albino rabbits (except for controls) were injected intravitreally with conjugate at various concentrations. Preoperative and postoperative ocular examinations and electroretinography were performed until animals were killed for histology. Part 2: Long-Evans pigmented rats had choroidal neovascularization (CNV) induced by argon laser. One eye per animal was injected with concentration 10M (safe dose), whereas the other eyes were controls and received 30 microL of sterile water at different time intervals after laser application. Fluorescein angiography was performed at various time points to evaluate the lesions and confirm presence of CNV. Animals were euthanized. The eyes were immediately enucleated and prepared for histologic examination. RESULTS: Part 1: No clinical changes were seen in groups receiving 10(-8)M, 10(-7)M, 10(-6)M, and 10(-5) M of conjugate. Electroretinography showed decreasing b-wave amplitude in groups receiving 10(-4) M and 10(-3) M; cataracts also developed in these eyes. Part 2: Fluorescein angiography revealed that intravitreal injection of somatostatin conjugate JF-10-81 favorably affected the development of CNV when the treatment was performed at least 1 week after the laser application. These results were statistically significant. Histologic analysis results of eyes treated 2 weeks after laser application also showed significant benefit. CONCLUSIONS: Part 1: Camptothecin-somatostatin conjugate injected intravitreally appeared safe at concentrations of 10(-5)M or less. Part 2: Conjugate JF-10-81 at a concentration of 10(-5)M administered intravitreally 1 to 3 weeks after laser demonstrated statistically significant efficacy in the treatment of choroidal neovascularization.

Photoablative inlay laser in situ keratomileusis (PAI-LASIK) in the rabbit model.

PURPOSE: To evaluate the suitability, biocompatibility, and efficacy of a proprietary hydrogel photoablative inlay (PAI) for use during laser in situ keratomileusis (LASIK). SETTING: Laboratory study, Tulane University Health Sciences Center, New Orleans, Louisiana, USA. METHODS: Eight rabbits (1 eye each) underwent the PAI-LASIK procedure; 4 eyes had a disk-shaped inlay and 4, a donut-shaped inlay. Preoperatively, the hydrogel material was ablated with a programmed correction of 5.0 diopters of hyperopia or myopia. RESULTS: The eyes were followed for 1 to 16 months. No eye showed signs of rejection or extrusion of the PAI. There was no significant difference in corneal clarity or the healing rate between eyes with donut-shaped PAIs and those with disk-shaped PAIs. One eye with a donut-shaped PAI had minimal corneal haze. The remaining inlays did not opacify or fracture during ablation. CONCLUSION: The hydrogel material can be used for the proposed PAI-LASIK procedure.

The effect of indocyanine green pretreatment on the parameters of transscleral diode laser thermotherapy-induced threshold coagulation of the ciliary body.

BACKGROUND AND OBJECTIVE: To study the effect of indocyanine green (ICG) pretreatment on threshold parameters of transscleral diode laser thermotherapy-induced threshold coagulation of the ciliary body. The procedure was termed 'cyclothermotherapy' based on the long duration (15-60 seconds) of diode laser application. STUDY DESIGN/MATERIALS AND METHODS: The right eyes of nine young adult New Zealand white rabbits underwent transscleral cyclothermotherapy (TCT, Group 1), TCT following ICG pretreatment (Group 2), and external manipulation of the ciliary body alone (Group 3). Rabbits were sacrificed after 24 hours; specimens were evaluated with gross examination and light microscopy. RESULTS: Thresholds were 30 J/cm2 (TCT) and 4.5 J/cm2 (TCT with ICG). Widespread structural damage was seen in the ciliary processes and the ciliary body in Groups 1 and 2. In Group 3, external manipulation of the ciliary body caused hemorrhage and structural damage confined to the ciliary processes. CONCLUSION: ICG pretreatment reduced the energy necessary to cause a threshold lesion with TCT in nonpigmented rabbits.

The role of recombinant lysine-plasminogen and recombinant urokinase and sulfur hexafluoride combination in inducing posterior vitreous detachment.

PURPOSE: To determine the optimal method of generating plasmin in vitreous using recombinant lysine-plasminogen and recombinant urokinase and to determine its efficacy in inducing posterior vitreous detachment when combined with sulfur hexafluoride. METHODS: Plasmin concentration of the rabbit vitreous after separate and combined intravitreal administrations of recombinant lysine-plasminogen and recombinant urokinase was tested in 78 rabbits to determine the optimal method of administration. The safety and efficacy of these agents and sulfur hexafluoride in inducing complete posterior vitreous detachment (total separation of the vitreous apart from vitreous base) were also evaluated. RESULTS: The highest plasmin concentration in vitreous was measured 10 minutes after injection. Intravitreal administration of recombinant lysine-plasminogen and recombinant urokinase did not cause any toxicity findings up to concentrations of 100 microg and 200 IU, respectively, on funduscopy, electroretinography, and histopathologic studies. When combined with sulfur hexafluoride injection, separate intravitreal administrations of 75 microg/0.1 mL of recombinant lysine-plasminogen and 15 IU/0.1 mL of recombinant urokinase induced complete posterior vitreous detachment in 75% of the eyes compared with 13% in eyes that received sulfur hexafluoride injection combined with balanced salt solution, recombinant lysine-plasminogen, or recombinant urokinase. CONCLUSIONS: Plasmin was effectively generated in the vitreous after separate intravitreal administrations of recombinant lysine-plasminogen and recombinant urokinase. When combined with intravitreal gas injection, this method of plasmin production induced complete posterior vitreous detachment in 75% of the eyes.

The effect of intraoperative retinal manipulation on the underlying retinal pigment epithelium: an experimental study.

PURPOSE: Retinal pigment epithelial changes described after vitreoretinal surgery may result from localized compression injury caused by intentional or inadvertent contact with vitreoretinal instruments. The authors evaluated these changes resulting from manipulation of the retina without frank retinal injury. METHODS: One eye each of six pigmented rabbits underwent surgery during which the inner retinal surface was touched at several points with a 20-gauge silicone-tipped subretinal fluid cannula without causing a retinal break or subretinal hemorrhage. The rabbits were followed-up with indirect ophthalmoscopy, fundus photography, and fluorescein angiography, and were killed at 1 hour, 1 week, or 2 weeks. Light microscopy was used for histopathologic evaluation. RESULTS: On fluorescein angiography, diffuse leakage noted at the injury sites significantly decreased by the fourth day and almost completely disappeared by the second week. Disruption of photoreceptor outer segments and retinal pigment epithelium in the early specimens, and irregular pigmentation, proliferation, and migration of the retinal pigment epithelium at 1 and 2 weeks were the prominent features on histopathologic examination. CONCLUSIONS: Intraoperative manipulation of the attached retina may cause significant pigment epithelium displacement and proliferation and varying degrees of disorganization of normal retinal architecture in the absence of clinically evident retinal breaks and subretinal hemorrhages.

Transpupillary thermotherapy threshold parameters: funduscopic, angiographic, and histologic findings in pigmented and nonpigmented rabbits.

PURPOSE: To evaluate the effect of pigmentation on threshold fluence levels, needed to produce visible and angiographic lesions, of transpupillary thermotherapy (TTT) in rabbits. METHODS: Six pigmented and nine nonpigmented rabbits underwent TTT with an 810-nm diode laser coupled to a slit-lamp biomicroscope using a spot size of 2 or 3 mm. The power ranged from 80 to 200 mW with 1 to 3 minutes of laser exposure for pigmented rabbits and 750 to 1800 mW with 1 minute of exposure for albino rabbits. These parameters were also evaluated after compression of the globe using the contact lens to induce blanching of the optic nerve head. After the experiment, the eyes were enucleated under deep anesthesia, and the animals were killed immediately. RESULTS: In pigmented rabbits, the threshold fluence with the 2-mm spot size was 229 J/cm2 without compression and 153 J/cm2 with compression. With the 3-mm spot size, the threshold decreased from 200 to 150 mW as the duration of exposure lengthened (2 or 3 minutes), increasing the fluence from 170 to 382 J/cm2. In nonpigmented rabbits, the threshold fluence with the 2-mm spot size was 2,865 J/cm2 without compression and 2,674 J/cm2 with compression. With the 3-mm spot size, the threshold fluence of 1,528 J/cm2 was not affected by compression. Histopathologic studies showed transretinal damage at the lowest levels necessary to achieve angiographic evidence of a treatment lesion or a barely visible funduscopic lesion at the time of treatment. CONCLUSIONS: Nonpigmented rabbits required more than a 12-fold increase in total TTT fluence compared with pigmented rabbits with the 2-mm spot size and a ninefold increase with the 3-mm spot size. Inner and outer retinal damage was seen histopathologically at these levels.

Transpupillary thermotherapy threshold parameters: effect of indocyanine green pretreatment.

PURPOSE: To evaluate the effect of combined treatment with systemic indocyanine green (ICG) on threshold fluence levels of transpupillary thermotherapy (TTT) in rabbits. METHODS: Four pigmented rabbits and 13 nonpigmented rabbits were studied. TTT was performed on normal rabbit choriocapillaris using an 810-nm diode laser via slit-lamp biomicroscope delivery through a Goldmann macular lens. Laser spot size, power, and duration of laser exposure were varied to achieve a range of TTT fluences for threshold testing in both albino and pigmented rabbit fundi. Intravenous ICG pretreatment at doses of 0.41 to 10 mg/kg was initiated at varying times before TTT treatment. After the experiment, the eyes were enucleated under deep anesthesia, the animals were killed, and the eyes were prepared for light microscopy. RESULTS: When intravenous ICG pretreatment was employed, there was a dose-dependent decrease in the TTT fluence threshold as compared with known threshold values. At threshold fluences, histopathologic sections revealed damage to all layers of the retina in addition to choriocapillaris damage. CONCLUSION: Intravenous ICG pretreatment can be used to lower the TTT threshold fluence and irradiance required to create angiographically visible lesions in the normal rabbit choriocapillaris. Damage was seen in all layers of the retina and choriocapillaris at threshold levels when TTT was used alone or in combination with ICG pretreatment.

The role of scleral buckle in experimental posterior penetrating eye injury.

PURPOSE: Although episcleral buckles are frequently placed as an additional procedure to vitreoretinal surgery, little is known about their independent effect after ocular trauma. The authors created a posterior penetrating ocular injury model to evaluate the isolated role of primary episcleral buckle placement. METHODS: Twenty eyes underwent surgery. The penetrating injury consisted of two 5-mm circumferential incisions placed five clock hours apart and 8 mm behind the limbus. A segmental episcleral buckle was placed over a randomly chosen injury site after wound closure. The degrees of fibrous proliferation, traction, and the presence of retinal detachment were evaluated on follow-up examinations. After enucleation and initial fixation, tissue sectioning was performed, and the greatest dimension of the fibrous proliferation at both wound sites was measured with a caliper. RESULTS: Two eyes were excluded from the study. Three eyes developed a retinal detachment; the remaining 15 eyes showed varying degrees of proliferation and traction on the retina. The greatest dimension of the fibrous proliferation at the buckle site (1.22 +/- 1 mm) was significantly different from that at the nonbuckle site (2 +/- 1.45 mm, P = 0.01). CONCLUSIONS: Primary episcleral buckle placement at the time of surgical repair reduces vitreous traction from the buckle site and decreases the degree of fibrovascular proliferation.