Laser-induced intrachoroidal dexamethasone drug delivery system to posterior eye segment.

PURPOSE: To investigate the feasibility of laser-induced intrachoroidal dexamethasone (DEX) delivery as a potentially useful therapy for adjusting the most effective drug level to the posterior segment eye diseases. METHODS: An implant was prepared by dissolving poly(DL-lactide) and DEX. In vitro release of DEX was evaluated at 7, 14, and 28 days by ELISA. In vivo, a DEX implant was inserted into a rabbit choroid, and 10, 50, or 200 burns of photocoagulation were applied at the implant lesion. After treatment, the vitreous humor was immediately aspirated and the DEX level was measured by liquid chromatography/mass spectrometry/mass spectrometry. Furthermore, the vitreous DEX level was measured at 1, 7, 14, and 28 days after implantation and 50 burns of photocoagulation. The toxicity of the laser-induced DEX implant was evaluated by ophthalmoscopy and light microscopy. Endotoxin-induced uveitis (EIU) was induced after DEX implantation and photocoagulation, and anti-inflammatory activities were evaluated by grading clinical signs, protein concentrations, and histopathologic studies. RESULTS: Photocoagulation significantly increased the DEX release from the implant at 7 days in vitro. In vivo, the DEX implant exposed to 10, 50, and 200 burns of photocoagulation increased the vitreous DEX levels in a dose-dependent manner. The vitreous DEX level in the DEX implant applied to 50 burns of photocoagulation peaked 1 day after treatment. The laser-induced DEX implant showed no retinal abnormalities except the implantation site, and significantly inhibited the EIU. CONCLUSIONS: Laser-induced intrachoroidal DEX delivery controls the DEX level in the vitreous humor and effectively prevents the experimental uveitis.

Inhibition of ocular angiogenesis by diced small interfering RNAs (siRNAs) specific to vascular endothelial growth factor (VEGF).

The effects of diced small interfering RNAs (siRNAs) designed for vascular endothelial growth factor (VEGF) on the expression of VEGF in human retinal pigment epithelial cell line ARPE-19 cells in vitro and on corneal angiogenesis in vivo were examined. The exposure to diced siRNAs significantly reduced the VEGF mRNA expression in ARPE-19 cells with minimal toxicity. In suture-induced corneal angiogenesis models, diced siRNAs minimized the severity of angiogenesis. Histological analysis displayed no particular tissue damage in the conjunctiva where siRNA was injected. The approach using diced siRNAs can be a new tool for various neovascular ocular diseases.

Inhibitory effect of triamcinolone acetonide on corneal neovascularization.

BACKGROUND: Corneal neovascularization (NV) plays an important role in the pathogenesis of corneal disorders. Recently, triamcinolone acetonide (TA) has been reported as a potential treatment for ocular angiogenesis. However, there are no reports on the inhibitory effect of TA on the corneal NV. METHODS: Triamcinolone acetonide (2 mg) was administered to four rabbits' eyes by a subconjunctival injection immediately after a basic fibroblast growth factor (bFGF)-pellet was placed into the cornea. As a control, four eyes received an injection of distilled water. Four weeks later, the inhibition of corneal NV was evaluated as the percentage ratio of the vessel invasion area to the area that was sandwiched between the pellet and the limbus cornea. To identify the characteristic appearance of new corneal vessels, the control cornea was examined by using the antibody of vascular endothelial growth factor (VEGF). To confirm TA concentration in TA-treated corneas, the TA level was measured using high-performance liquid chromatography. RESULTS: Neovascularization from the limbus to the pellet was detected in control eyes 4 weeks after the bFGF pellet implantation. TA-treated eyes demonstrated the inhibition of the neovascular response to the pellet. The severity of NV as compared between control and TA-treated eyes was statistically significant (P<0.05). Morphologically, new vessel growth was shown in the control cornea, and endothelial cells of new vessels were positively stained with the antibody of VEGF. TA concentration in TA-treated corneas at 2 weeks showed 63.5+/-42.8 microg/g (n=4, mean +/- SD), while TA was not detected in control and TA-treated corneas at 4 weeks. The level of TA was effectively maintained for at least 2 weeks after the subconjunctival injection. CONCLUSION: We have demonstrated that subconjunctival TA administration inhibited rabbit corneal NV. This agent may prove useful in the treatment of corneal angiogenic disorders.

Hyaluronan synthases, hyaluronan and its CD44 receptors in the posterior segment of rabbit eye.

To understand the possible roles of the hyaluronan synthetase (HAS)/hyaluronan (HA)/CD44 signaling system in the posterior eye segment, we investigated the expression of rabbit HAS isoforms and CD44 mRNA by RT-PCR and the level of HA by using HA assay and immunohistochemistry. HA was detectable in vitreous, retina and choroid. The expression of three HAS isoforms was clearly detected in both retina and choroids. Rabbit choroid showed a significant increase of the HAS2 and HAS3 expression compared with rabbit retina (HAS2 p = 0.0014 < 0.05; HAS3 p = 0.0006 < 0.05). Similarly, mRNA expression of CD44 was detected in both retina and choroids. This evidence may suggest that the HAS/HA/CD44 signaling system is important in maintaining the functional structure of retina and choroid.

The effect of indocyanine green on cultured retinal glial cells.

BACKGROUND: Recently, indocyanine green (ICG) has been utilized to visualize inner limiting membrane in vitreous surgery. However, the safety of ICG injected into the vitreous has not been well established. The possible toxicity of ICG on Muller cells was investigated using cultured rat retinal glial cells (RGCs). METHODS: Rat RGCs were cultured in Dulbecco modified Eagle medium supplemented with 20% fetal calf serum. The cytotoxicity of ICG was assayed with viable cell number and resazurin metabolic assay. The expression of the apoptosis-related gene bcl-2 was examined with real-time polymerase chain reaction analysis. RESULTS: The effects of ICG on the viability of rat RGCs were tested at two different concentrations (0.05% and 0.5%). ICG significantly decreased the viable cell number of RGCs at 0.5%, while there was no significant effect at 0.05%. Similarly, the metabolic activity to resazurin was significantly decreased by exposure to 0.5% ICG. However, ICG showed little effects on resazurin metabolism at 0.05%. The expression levels of bcl-2 mRNA were higher in cells treated with 0.5% ICG than in those treated with 0.05% ICG and untreated control cells. CONCLUSION: The data suggest that ICG initiates the death of RGCs at high concentrations, in part, through apoptosis-related signal pathways.