[A rabbit model for human cytomegalovirus retinitis].

PURPOSE: To develop a rabbit model for human cytomegalovirus(HCMV) retinitis. METHODS: 0.1 ml of 1 x 10(6) plaque forming units/ml HCMV was injected into the vitreous cavity of 10 pigmented rabbit eyes. The eyes were examined ophthalmoscopically on days 1, 2, 3, 4 and 7 and once a week thereafter until 4 weeks after inoculation. Vitreal and retinal findings were graded from 0+ to 4+ on a scale of increasing severity. In addition, we examined the enucleated eyes 3 weeks after HCMV inoculation by histological and immunohistochemical techniques. RESULTS: All injected eyes developed vitreoretinal lesions. Vitreous opacities appeared the next day and increased until 4 days after HCMV inoculation. Whitish retinal exudates occurred on day 3 and increased until 3 weeks after HCMV inoculation. Vitreoretinal lesions then disappeared by 4 weeks after inoculation. Histological examination revealed intraretinal infiltration of inflammatory cells and disorganization of the inner retinal architecture. HCMV antigens were detected inside the retina by immunofluorescence using anti early protein antibody against HCMV. CONCLUSIONS: The results indicate that this rabbit model can be useful to develop and evaluate a new treatment modality for cytomegalovirus retinitis.

Scleral plug of biodegradable polymers containing ganciclovir for experimental cytomegalovirus retinitis.

PURPOSE: To evaluate the efficacy of a biodegradable scleral plug containing ganciclovir (GCV) in a rabbit model of human cytomegalovirus (HCMV) retinitis. METHODS: To develop a rabbit model for HCMV retinitis, HCMV solution was injected once into the vitreous cavity of pigmented rabbits. The treated animals were divided into three groups: group A received no treatment, group B was treated once with GCV solution, and group C was treated with a scleral plug containing GCV. Rabbits in group B received an intravitreal injection of GCV solution 1 week after HCMV inoculation. In group C, the scleral plug containing GCV was implanted in the vitreous of the rabbits 1 week after HCMV inoculation. Ophthalmoscopically, vitreoretinal findings in each group were graded from 0+ to 4+ every week for 4 weeks after HCMV injection. RESULTS: Eyes of group A rabbits showed whitish retinal exudates and vitreous opacities 3 days after HCMV inoculation. These materials increased gradually until 3 weeks after HCMV inoculation. Scores for vitreoretinal lesions were significantly lower in eyes of group B rabbits compared with those of group A at 1 week after GCV injection (P < 0.05). However, vitreoretinal inflammation in eyes of group B rabbits increased again thereafter, and no significant difference in inflammation between groups A and B was found 2 weeks after GCV injection. In eyes of group C, scores for vitreoretinal lesions were significantly lower compared with those in both group A and group B at 3 weeks after HCMV inoculation (P < 0.01). CONCLUSIONS: The results demonstrated that sustained release of GCV into the vitreous cavity with biodegradable scleral plugs was effective for the treatment of experimentally induced HCMV retinitis in rabbits.

Effect of particle size of polymeric nanospheres on intravitreal kinetics.

In this study, we injected nanospheres containing a fluorescein derivative into the vitreous cavity of pigmented rabbit eyes and evaluated their intraocular kinetics as drug carriers in vivo. Polystyrene nanospheres (2 microm, 200 nm and 50 nm in diameter) containing a fluorescein derivative were used in this study. A suspension of each particle was prepared by diluting with distilled water at a concentration of 10 microg/ml equivalent to sodium fluorescein. The suspension of nanospheres was injected once into the vitreous cavity of unilateral eyes of pigmented rabbits. A sodium fluorescein solution of the same concentration was injected once into the vitreous cavity of the other eye as the control. The intraocular kinetics of nanospheres was evaluated by measuring vitreous fluorescence using a scanning fluorophotometer. To investigate elimination pathways of nanospheres in detail, serial cross-sections of the eyes were examined with a fluorescence microscope. The fluorescence derived from nanospheres was observed in the vitreous cavity for over 1 month (2 microm: t(1/2) = 5.4 +/- 0.8 days, 200 nm: t(1/2) = 8.6 +/- 0.7 days, 50 nm: t(1/2) = 10.1 +/- 1.8 days), whereas that in the control eyes completely disappeared within 3 days (t(1/2) = 7.8 +/- 0.7 h). The elimination half-life from the vitreous cavity correlated well with the particle diameter (r = -0.997, p = 0.007). Histological studies using a fluorescence microscope revealed that nanospheres with a diameter of 2 microm were seen in the vitreous cavity and trabecular meshwork, while nanospheres with a diameter of smaller than 200 nm were also observed in the retina as well as these tissues. Our findings indicated that nanospheres may be beneficial as a drug carrier to the retina, vitreous and trabecular meshwork.

Long-term sustained release of ganciclovir from biodegradable scleral implant for the treatment of cytomegalovirus retinitis.

The previous scleral implant composed of poly(D, L-lactide-co-glycolide) with ganciclovir (GCV) had some disadvantages such as the second burst in the late phase of release. In this study, the GCV release rate from scleral implants was modified by blending poly(D,L-lactide) (PLA) of two different molecular weights. The scleral implants were prepared by blending PLA-70000 (molecular weight: 70000) and PLA-5000 (molecular weight: 5000) or PLA-130000 (molecular weight: 130000) and PLA-5000 at weight ratios of 100/0, 95/5, 90/10, 80/20, and 0/100. In vitro release tests were performed in 0.1 M phosphate-buffered solution (pH 7.4) at 37 degrees C. An increase in the blended amount of PLA-5000 clearly accelerated the GCV release and the onset of the second burst in the late phase of release tended to delay. The two implants both prepared at a blend ratio of 80/20 successfully prevented the second burst and the GCV release profiles followed the pseudozero-order kinetics after the initial burst as resulting from a diffusional mechanism following Higuchi's equation. Duration of the sustained GCV release could be controlled by changing the blending ratio of high and low molecular weight PLA. The 25% GCV-loaded scleral implants composed of PLA-70000 and PLA-5000 with a blending ratio of 80/20 were implanted in pigmented rabbit eyes. The GCV concentrations in the vitreous after implantation of PLA-70000/PLA-5000 scleral implant with a blending ratio of 80/20 were maintained in the range of effective level for 6 months without a significant burst. Our results suggest that the blended implants are promising for the intraocular controlled drug delivery over a period of several months to one year to treat cytomegalovirus retinitis.

Biodegradable scleral implant for controlled intraocular delivery of betamethasone phosphate.

We evaluated nail-like, biodegradable scleral implants as a controlled intraocular delivery system of betamethasone phosphate (BP) for the treatment of chronic uveitis using pigmented rabbits. The scleral implants, which contained 10% and 25% of BP (weight 8.5 mg; length 5 mm), were made of poly (D,L-lactide-co-glycolide) (PLGA). In vivo release and retinal toxicity after implantation were also studied in pigmented rabbits. The in vitro release studies demonstrated the 10% and 25% BP-loaded scleral implants released BP in a biphasic release pattern for at least 1 month. The BP concentrations in the vitreous and the retina/choroid after application of scleral implants in pigmented rabbit eyes stayed within the concentration range capable of suppressing inflammatory responses for over 1 month. The BP concentration was greater in the retina/choroid than in the vitreous throughout the study. No substantial toxic reactions in the retina were observed by electroretinography. Our findings suggest that the BP-loaded scleral implant may be a promising device for treatment of chronic uveitis.

Biodegradable scleral implant for intravitreal controlled release of ganciclovir.

BACKGROUND: The aims of this study were to develop biodegradable scleral implants that could overcome previously reported disadvantages such as an adverse burst in the late phase of release and to investigate the release profile of modified scleral implants in vitro and in vivo. METHODS: The modified scleral implants (weight 8.5 mg, length 5 mm) were made of mixtures of poly(DL-lactide) (PLA) with different molecular weights and contained 25 weight % of ganciclovir (GCV). The release of GCV was evaluated in vitro by spectrophotometry. Intravitreal GCV concentrations in vivo were measured by high-performance liquid chromatography following plug implantation in pigmented rabbits. The biocompatibility of the device was determined by indirect ophthalmoscopy and light microscopy. RESULTS: The in vitro release studies showed stable, long-term sustained and slow release. The in vivo release studies showed that the implants had long-term release in the diffusional phase of the triphasic release pattern and only a minor adverse burst of GCV in the late phase. No significant retinal toxicity was observed by histologic examination. CONCLUSION: Our findings showed that this newly modified scleral implant may provide suitable intravitreal drug delivery for treatment of cytomegalovirus retinitis.

Biodegradable scleral implant for intravitreal controlled release of fluconazole.

PURPOSE: To evaluate the feasibility of using a biodegradable polymeric scleral implant containing fluconazole (FLCZ), a bis-triazole antifungal agent, as a potential intravitreal-controlled drug delivery system. METHODS: The scleral implants, loaded with 10, 20, 30, and 50% FLCZ, were prepared with biodegradable polymers of poly (DL-lactide-co-glycolide). Those with all loading doses were used for the in vitro release studies; those with 30% FLCZ were used for the intravitreal release studies in pigmented rabbits. The in vitro and in vivo release rates of FLCZ from the implants were measured periodically with spectrophotometry and high performance liquid chromatography, respectively. The effects of the implants on ocular tissues were evaluated ophthalmoscopically, histologically, and electrophysiologically. RESULTS: The scleral implants loaded with 10, 20, and 30% doses gradually released FLCZ over 4 weeks in vitro; those with 50% FLCZ released most of the drug in one week. FLCZ concentration in the rabbit vitreous remained within the 99% inhibitory concentration for Candida albicans for 3 weeks after implantation. The scleral implant gradually biodegraded, and it disappeared by 4 months after implantation. The electrophysiologic and histopathologic findings demonstrated no substantial toxic reactions in the ocular tissues. CONCLUSION: The current study suggests that a biodegradable, polymeric scleral implant containing FLCZ may be a promising intravitreal drug delivery system to treat fungal endophthalmitis.

Biodegradable polymeric device for sustained intravitreal release of ganciclovir in rabbits.

PURPOSE: A scleral plug made of biodegradable polymer implanted at the pars plana was evaluated to determine its ability to control the intravitreal release of ganciclovir. METHODS: Scleral plugs containing 25% ganciclovir were prepared with poly(lactic-glycolic acid) (molecular weight, 121 kDa). The release of ganciclovir was evaluated in vitro by spectrophotometry. In vivo intravitreal ganciclovir concentrations were measured by high performance liquid chromatography following plug implantation in pigmented rabbits. The biocompatibility of the device was determined by indirect ophthalmoscopy, electroretinography, and light and electron microscopy. RESULTS: The in vitro study showed that the plug released ganciclovir throughout a 10-week period. The in vivo study demonstrated that the plugs maintained the drug concentration in the vitreous in a therapeutic range adequate to treat cytomegalovirus (CMV) retinitis for 12 weeks. No significant retinal toxicity was observed. CONCLUSIONS: This study demonstrated that this drug delivery system can potentially be useful to treat CMV retinitis.

Implantable biodegradable polymeric device in the treatment of experimental proliferative vitreoretinopathy.

We investigated the use of a scleral plug of biodegradable polymer implanted at the pars plana to create a controlled drug-delivery system in the vitreous. We evaluated the efficacy of a plug containing doxorubicin hydrochloride to treat experimental proliferative vitreoretinopathy (PVR) in pigmented rabbits. An implantable device on the sclera, which imitates a scleral plug, containing 1% doxorubicin, was prepared with poly(lactic acid) (molecular weight, 20,000). The release of doxorubicin in phosphate-buffered saline was evaluated by spectro-photometry. After pars plana vitrectomy and plug implantation, concentrations of doxorubicin in the vitreous humor of the rabbits were measured by high performance liquid chromatography. The release profiles were evaluated during 5 weeks in vitro and 4 weeks in vivo. Cultured homologous fibroblasts were injected into the vitreous space to induce experimental PVR after gas compression of the vitreous. The scleral plugs were implanted at the pars plana in treatment animals (n = 11). Control rabbits (n = 11) were followed up without implantation after PVR induction. All eyes of the control group developed tractional retinal detachment at day 28, while the incidence of retinal detachment was decreased to 64% in the treated eyes. (P = 0.002). The implantation of the scleral plug effectively inhibited intravitreous proliferation of fibroblasts. This study demonstrated that the scleral plug of biodegradable polymers may have potential as a treatment modality for PVR.

In vitro drug release behavior of D,L-lactide/glycolide copolymer (PLGA) nanospheres with nafarelin acetate prepared by a novel spontaneous emulsification solvent diffusion method.

Nanospheres with D,L-lactide/glycolide copolymer (PLGA) were prepared as a biodegradable and biocompatible polymeric carrier for peptide drugs by a novel spontaneous emulsification solvent diffusion method. Nafarelin acetate (NA), a luteinizing hormone-releasing hormone analogue, was employed as a model peptide drug to investigate the encapsulation efficiency. The drug and PLGA, dissolved in an acetone-dichloromethane-water mixture, were poured into an aqueous solution of polyvinyl alcohol under moderate stirring at room temperature. Spontaneous emulsification arising from a rapid diffusion of acetone from the organic to the aqueous phase enables preparation of PLGA submicron spheres 200-300 nm in size. The entrapment of NA in nanospheres was improved by blending low molecular weight (Mw = 4500) PLGA with higher molecular weight PLGA due to the synergistic effect of the rapid deposition of PLGA and the ionic interaction between NA and PLGA. By coadmixing a small amount of negatively charged phospholipids such as dipalmitoyl phosphatidylglycerol or dicetyl phosphate, the leakage of water-soluble NA was further prevented. The NA encapsulated in PLGA nanospheres was more stable than native NA in acidic medium (pH = 1.2). The drug-release behavior from nanospheres suspended in the disintegration test solution no. 1 (Japanese Pharmacopeia XII) exhibited a biphasic pattern. It was found tht the initial burst of release might be due to the degradation of the PLGA chain, as monitored by gel permeation chromatography. At a later stage, the drug was released more slowly, the rate of which was determined by the diffusion of the drug in the porous matrix structure.(ABSTRACT TRUNCATED AT 250 WORDS)