Vitreous Cavity Of Rabbit Eyes Research Articles

An in situ-forming polyzwitterion hydrogel: Towards vitreous substitute application

Development of a biostable and biosafe vitreous substitute is highly desirable, but remains a grand challenge. Herein, we propose a novel strategy for constructing a readily administered vitreous substitute based on a thiol-acrylate clickable polyzwitterion macromonomer. A biocompatible multivinyl polycarboxybetaine (PCB-OAA) macromonomer is designed and synthesized, and mixed with dithiothreitol (DTT) via a Michael addition reaction to form a hydrogel in vitreous cavity. This resultant PCB-OAA hydrogel exhibits controllable gelation time, super anti-fouling ability against proteins and cells, excellent biocompatibility, and approximate key parameters to human vitreous body including equilibrium water content, density, optical properties, modulus. Remarkably, outperforming clinically used silicone oil in biocompatibility, this rapidly formed hydrogel in the vitreous cavity of rabbit eyes remains stable in vitreous cavity, showing an appealing ability to prevent significantly inflammatory response, fibrosis and complications such as raised intraocular pressure (IOP), and cataract formation. This zwitterionic polymer hydrogel holds great potential as a vitreous substitute.

Intravitreal bromfenac liposomal suspension (100 μg / 0.1 ml). A safety study in rabbit eyes

IntroductionThere is a need to find alternative treatments for MEe. Bromfenac has shown promise in inhibiting the COX-2 enzymatic pathway that partially causes the inflammatory cascade which contributes to the precipitation of ME. However, like other NSAID's, its intraocular half-life is limited. We hypothesize that a delayed-release liposome formulation containing bromfenac might provide a similar anti-inflammatory effect as long-lasting steroid release systems without the well-known steroidal side-effects. We introduced a novel formulation with these characteristics into the vitreous cavity of rabbit eyes in order to evaluate its safety profile. Material and methods10 left eyes of rabbits were injected with the liposome-encapsulated bromfenac suspension (100 μg/0.1 ml). Basal ERG's were recorded. Total follow-up time was 3 months, at which point ERG's were repeated and eyes were enucleated for histopathological study. Total amplitude and implicit times were recorded. A difference of 25% in either recording was considered significant. Significance was assessed using the paired-t test and Wilcoxon matched-pairs signed-rank test. A p-value of <0.05 was considered significant. ResultsNo significant changes were recorded in ERG measurements after 3 months when compared to basal measurements. Histopathological analysis of retinal specimens found no traces of liposome-induced toxicity. ConclusionThe liposome-encapsulated bromfenac suspension (100 μg/0.1 ml) is not toxic and has been proven safe to use in an animal model. Therefore, this formulation shows promise as a possible future alternative treatment for ME and should be further studied to show its biological effect and efficacy.

Real-Time Intraocular Pressure Measurements in the Vitreous Chamber of Rabbit Eyes During Small Incision Lenticule Extraction (SMILE)

ABSTRACTPurpose: To investigate real-time intraocular pressure (IOP) during small incision lenticule extraction (SMILE) in rabbit eyes for myopia correction.Methods: During SMILE, real-time IOP was measured in the vitreous cavity of rabbit eyes with an optic fiber pressure sensor (OFPS). Two groups (n = 6 for each) underwent surgery, one group for a −2.00 diopter (D) refractive spherical correction and the other for a −6.00 D correction.Results: During surgery, the IOP increased once the glass contact attached to the cornea (Pre-suction), and peaked 83.94 mmHg (SD ± 23.87 mmHg) for the −2.00 D group and 89.17 mmHg (SD ± 22.66 mmHg) for the −6.00 D group, both average values were less than 110 mmHg when suction was initiated to fix the glass contact onto the cornea (Suction on). It then fell to 74.81 mmHg (SD ± 20.64 mmHg) and 76.94 mmHg (SD ± 27.43 mmHg), respectively, and remained stable during lenticule creation (Cutting). After suction stopped (Suction off), IOP fell steeply. During lenticule separation/extraction, the change in IOP was 32.26 mmHg (SD ± 2.91 mmHg). Notably, the average duration of elevated IOP during the surgery was 166.05 s (no longer than 3 min).Conclusions: The IOP fluctuations in the vitreous cavity using an OFPS in a rabbit model during SMILE showed that real-time IOP significantly was increased during Pre-suction, Suction on, Cutting, Suction off, and lenticule separation/extraction compared to baseline IOP, although, peaked at Suction on. Neither the degree of myopic correction nor central corneal thickness significantly affected these changes in IOP.

Biocompatibility and retinal support of a foldable capsular vitreous body injected with saline or silicone oil implanted in rabbit eyes

The aim of this study was to evaluate over a 180-day period the biocompatibility and retinal support of a foldable capsular vitreous body injected with either saline or silicone oil implanted in rabbit eyes. A standard three-port pars plana vitrectomy was performed, and foldable capsular vitreous bodies were implanted into the vitreous cavity of rabbit eyes (n = 18). Silicone oil tamponade was used as the control group (n = 5). Of the foldable capsular vitreous body-implanted eyes, either saline (n = 9) or silicone oil (n = 9) was injected into the foldable capsular vitreous body to support the retina. The treated eyes were examined using a slit lamp with a non-contact slit-lamp lens, a tonopen, a non-contact specular microscope and a B-scan ultrasound during the 180-day implantation period. A histological examination was performed at 90 and 180 days. During the 180-day implantation period, no significant corneal keratopathy or intraocular inflammation was noted, and the intraocular pressure (IOP) and corneal endothelial numbers remained steady among the three groups. B-scan ultrasonography showed a smoothly increased echogenicity in front of the retina in group of foldable capsular vitreous bodies injected with saline. Gross examination showed that the foldable capsular vitreous bodies injected with saline or silicone oil smoothly supported the retina. The saline or silicone oil inside the foldable capsular vitreous body was homogeneous, transparent and filled the foldable capsular vitreous body. Histological examination showed no obvious abnormality of the cornea, ciliary body or retina in the foldable capsular vitreous body-implanted eyes. These results suggest that foldable capsular vitreous bodies injected with either saline or silicone oil showed good biocompatibility and retinal support in rabbit eyes over a 180-day implantation time.

An Injectable Oxidated Hyaluronic Acid/Adipic Acid Dihydrazide Hydrogel as a Vitreous Substitute

Vitrectomy is a common procedure for treating ocular-related diseases. The surgery involves removing the vitreous humor from the center of the eye, and vitreous substitutes are needed to replace the vitreous humor after vitrectomy. In the present study, we developed a colorless, transparent and injectable hydrogel with appropriate refractive index as a vitreous substitute. The hydrogel is formed by oxidated hyaluronic acid (oxi-HA) cross-linked with adipic acid dihydrazide (ADH). Hyaluronic acid (HA) was oxidized by sodium periodate to create aldehyde functional groups, which could be cross-linked by ADH. The refractive index of this hydrogel ranged between 1.3420 and 1.3442, which is quite similar to human vitreous humor (1.3345). The degradation tests demonstrated that the hydrogel could maintain the gel matrix over 35 days, depending on the ADH concentration. In addition, the cytotoxicity was evaluated on retina pigmented epithelium (RPE) cells cultivated following the ISO standard (tests for in vitro cytotoxicity), and the hydrogel was found to be non-toxic. In a preliminary animal study, the oxi-HA/ADH hydrogel was injected into the vitreous cavity of rabbit eyes. The evaluations of slit-lamp observation, intraocular pressure, cornea thickness and histological examination showed no significant abnormal biological reactions for 3 weeks. This study suggests that the injectable oxi-HA/ADH hydrogel should be a potential vitreous substitute.

Injection Site and Pharmacokinetics After Intravitreal Injection of Immunoglobulin G

The aim of this study was to investigate whether the pharmacokinetics differ after immunoglobulin G (IgG) intravitreal injections are done in the different sites of the vitreous cavity of rabbit eyes. To examine the pharmacokinetics in the vitreous and the retina/choroid, fluorescein isothiocyanate-labeled IgG (5 μg/50 μL) was injected from the superior pars plana into the vitreous cavity of rabbit eyes. An original intravitreal injection guide was used to fix the tip of the injection needle, with the tip fixed in either the superior-anterior vitreous (superior group) or in the posterior vitreous (posterior group). At 1 h, 1, 4, and 7 days after injection, the eyes were enucleated and frozen. The vitreous was cut into superior, inferior, and posterior vitreous sections, whereas the retina/choroid was cut into superior, inferior, and posterior retina/choroid sections. The IgG concentrations in the vitreous and the retina/choroid sections were then determined. In the posterior vitreous, the IgG concentration in the posterior group was significantly higher than that in the superior group at 1 h after injection (P < 0.05). However, 1 day after injection, no significant differences were noted between the 2 groups. At 4 days after injection, the drug was diffusely spread in both groups. In the posterior retina/choroid, the IgG concentration was essentially the same regardless of the injection site or the amount of time after injection. The IgG concentration in the superior retina/choroid was significantly higher in the superior group than in the posterior group at 1 h and 1 day after injection (P < 0.05). There were no differences noted between the 2 groups for the IgG concentrations in the 3 sections at 4 and 7 days after injection. Intravitreally injected IgG remains in the area of the injection, with more than 1 day required for it to spread diffusely within the vitreous. In the posterior retina/choroid, results suggested that the concentration of IgG may be equal regardless of the injection site. In the superior retina/choroid area that was near the site of the injection, the concentration of drug tended to be higher.

Safety of In Vivo Pharmacologic Vitreolysis with Recombinant Microplasmin in Rabbit Eyes

To investigate the safety of intravitreal microplasmin in rabbits and to confirm previous findings of posterior vitreous detachment (PVD). Different doses of microplasmin, from 12.5 microg to 250 microg, in 0.1 mL balanced salt solution (BSS) were injected into the vitreous cavity of rabbit eyes to induce PVD. Fellow eyes were injected with the same volume of BSS. Slit-lamp biomicroscopy, ophthalmoscopic fundus examinations, A- and B-mode ultrasonography, and electroretinography were performed to assess the retina. Electroretinograms (ERGs) were recorded up to 90 days after injection. Morphologic alterations were assessed by light microscopy, scanning electron microscopy (SEM), and transmission (TEM) electron microscopy. A slight aqueous flare and cells were observed in the anterior chamber after microplasmin and BSS injection. A slight inflammatory reaction was also observed transiently in the vitreous cavity. In control eyes, B-mode ultrasonography and SEM examination demonstrated that PVD did not develop after BSS injection. Intravitreal injections of 125 microg or greater of microplasmin induced complete PVD with an internal limiting membrane (ILM) devoid of vitreous collagen fibrils. Eyes injected with 12.5 microg microplasmin had partial PVD, and SEM showed residual fibrils covering the ILM. In all eyes, there was a transient reduction in the a- and b-waves of the ERG on days 2 through 7. The ERGs showed less effect with < 250 microg microplasmin. Intravitreal injection of recombinant microplasmin in the rabbit induces no ERG or retinal ultrastructural abnormalities. Pharmacologic vitreolysis with this agent may be a useful adjunct to vitreous surgery and could be used to induce PVD without vitreous surgery.

Effects of intravitreal indocyanine green injection in rabbits.

To report the clinical, electrophysiologic, and histologic findings of different concentrations of indocyanine green (ICG) injected into the vitreous cavity of rabbit eyes. Forty-two rabbits underwent intravitreal injection of 0.1 mL of ICG in three different concentrations: 0.5 mg/mL (250 mOsm), 5 mg/mL (270 mOsm), and 25 mg/mL (170 mOsm). Fellow eyes were injected with 0.1 mL of balanced salt solution. Biomicroscopy, ophthalmoscopy, electroretinography, fluorescein angiography, and histologic evaluation were performed. Eyes injected with 0.5 mg/mL of ICG showed b-wave latency delay on the first day after injection. Eyes injected with 5 mg/mL of ICG showed b-wave latency delay and decreased b-wave amplitude on the first and seventh days after injection; delayed a-wave latency on the first day after injection was also observed. Eyes injected with 25 mg/mL of ICG showed b- and a-wave amplitude and latency abnormalities during the entire follow-up. Direct correlation of increasing retinal edema proportional to the progressively increasing ICG concentrations was shown on histologic evaluation. Intravitreal ICG injection in rabbit eyes may impair retinal function and morphology proportional to the progressively increasing ICG dosages.

Ocular toxicity study of trypan blue injected into the vitreous cavity of rabbit eyes.

To evaluate the ocular toxicity of trypan blue (TB) injected into the vitreous cavity of rabbit eyes. TB is a dye that could be useful for staining epiretinal membranes during vitrectomy surgery. Ten New Zealand White (NZW) rabbits underwent gas-compression vitrectomy. Rabbits were divided into three groups to receive injections of 0.1 ml basic salt solution, 0.1 ml of a 0.06% TB solution or 0.1 ml of a 0.2% TB solution. Ocular toxicity was assessed by slit-lamp biomicroscopy, ophthalmoscopy, electroretinography and histology. Transient posterior capsule opacification was noted in all animals. No significant reductions in a-wave or b-wave amplitudes were found in any of the animals. Light and electron microscopic examination of the inferior retina in the 0.2% TB-treated eyes showed damaged photoreceptors and marked disorganization. Immunohistochemical staining for rhodopsin was strongly reduced in those sections and staining for proliferation with Ki-67 was positive. No histological abnormalities were found in the upper retina of the 0.2% TB-treated eyes or in any part of the retina of the 0.06% TB-treated or control eyes. No histological abnormalities were found in any of the anterior chamber angle specimens. Although no signs of toxicity were found after the prolonged presence of TB at a concentration of 0.06% in the vitreous cavity of rabbit eyes, marked damage occurred in the lower retina of 0.2% TB-treated eyes. The short-term presence of TB at a concentration of 0.06% in the vitreous cavity is harmless to the rabbit eye but a higher concentration of TB could be unsafe.

Role of Neutrophils in Breakdown of the Blood-Retinal Barrier Following Intravitreal Injection of Platelet-activating Factor

Breakdown of the blood-retinal barrier occurs in inflammatory conditions and in ischemic retinal diseases such as diabetic retinopathy. Platelet-activating factor (PAF) is a potent inflammatory mediator which increases vascular permeability. The purpose of this study was to determine if intravitreally-injected PAF would cause breakdown of the blood-retinal barrier and, if so, by what mechanism. Fluorescein angiography was performed before and at 0·5, 1, 2, 3 and 4 hr after PAF injection into the vitreous cavity of rabbit eyes and the eyes were enucleated immediately for light and electron microscopy. Slow flowing thrombi were observed in all PAF-injected eyes. Complete vascular occlusion was observed in 10 of 16 eyes after 3 and 4 hr. There was no fluorescein leakage in any of eyes before or at 0·5 or 1 hr after PAF injection. Fourteen of 20 eyes had fluorescein leakage at 2, 3 and 4 hr after PAF injection. The extent of fluorescein leakage correlated with the degree of polymorphonuclear leukocyte (PMN) margination, disruption of the endothelial cell layer, infiltration into vascular walls and migration into the vitreous cavity. PMNs appeared to migrate by both intercellular and transcellular routes across the endothelium. Pretreatment of rabbits with a PAF inhibitor, BN52021, prevented most of the abnormal findings.

Ocular toxicity of daunomycin: effects of subdivided doses on the rabbit retina after vitreous gas compression.

Daunomycin is a potent antiproliferative agent which has been shown to prevent experimental proliferative vitreoretinopathy. However, toxic effects on the rabbit retina have been reported even for the lowest effective doses. In a previous report we demonstrated that subdivided doses rather than single doses of daunomycin improves the efficacy in prevention of experimental proliferative vitreoretinopathy. To evaluate whether dose subdivision would also have an alleviating effect on drug toxicity, we administered 15 nmol daunomycin in doses of 10 nmol and 5 nmol 4 h apart into the vitreous cavity of rabbit eyes which had previously undergone vitreous gas compression. All contralateral eyes received sham treatment. Simultaneous electroretinographic recordings from both eyes on days 0, 3, 7, and 14 demonstrated a significant b-wave decline in drug-exposed eyes. Morphological studies on these eyes revealed no retinal damage. Our findings suggest that dose subdivision does not eliminate the retinal toxicity of daunomycin.

Effects of Inflammation and Surgery on Amikacin Levels in the Vitreous Cavity

Intraocular injection of amikacin is increasingly used in the treatment of endophthalmitis. We injected 400 micrograms of amikacin into the vitreous cavity of rabbit eyes to study its pharmacokinetics. Phakic, aphakic, and aphakic vitrectomized eyes were injected, and inflamed eyes were compared to control eyes. Vitreous concentrations were determined at two, eight, 24, and 48 hours, and clearance rates were calculated. Amikacin is cleared considerably more quickly from aphakic (half-life, 14.3 hours) than phakic control eyes (half-life, 25.5 hours) and even more quickly from aphakic vitrectomized eyes (half-life, 7.0 hours). Inflammation substantially increased the rate of clearance in aphakic eyes. In inflamed aphakic and aphakic vitrectomized eyes, vitreous drug levels were equal to or below the minimal inhibitory concentration for most organisms considered sensitive to amikacin at 24 hours. Supplementation of intraocular antibiotics may therefore be required in clinical settings.

Sodium hyaluronate as a carrier for intravitreal gentamicin an experimental study

Fifty mcg of gentamicin was combined with saline or with 0.8% sodium hyaluronate and injected into the vitreous cavity of rabbit eyes with moderate to severe Staphylococcus aureus endophthalmitis. Endophthalmitis was controlled in 9 of 10 eyes. There was no evidence of toxicity with either treatment regimen. Although the clearance study demonstrated statistical differences at all time points studied, the half-lives of both treatment regimens were similar (3.3 h for aqueous gentamicin and 3.6 h for sodium hyaluronate/gentamicin). These results suggest that the vitreous played a role in keeping the aqueous gentamicin in the eye for a longer time, as similar half-lives were shown with both types of treatment. Thus, if a vitrectomy has to be done for the treatment of endophthalmitis, as much as possible of the vitreous should be left in situ to maintain the drug for longer periods in the eye. Also, if it is necessary to remove all vitreous during vitrectomy, it may be more effective to administer the drug with sodium hyaluronate so as to prolong its action inside the eye.

The Role of Cellular Proliferation in an Experimental Model of Massive Periretinal Proliferation

Transplantation of from 10,000 to 750,000 autologous and homologous dermal fibroblasts into the vitreous cavity of rabbit eyes resulted in identical clinical findings leading to massive periretinal proliferation. A dose-response relationship between the number of cells injected and the extent of retinal traction that developed was established. The second phase of the experiment evaluated the effects of formalin and cobalt irradiation on the fibroblasts' ability to cause traction retinal detachment. Cell proliferation or the attainment of a critical mass of living cells was necessary for the development of traction retinal detachment.

Intravitreal Autotransplantation of Fibroblasts

Tissue cultured skin fibroblasts autotransplanted into the vitreous cavity of rabbit eyes formed intravitreal strands that grew toward the medullary ray and optic nerve head and caused preretinal puckers and traction detachment. After four weeks, 32 of 51 eyes (63%) developed these changes. Light and electron microscopy revealed initial cell death. Remaining cells aligned themselves quickly into strands and began to multiply as shown by tritiated thymidine methyl incorporation. The appearance of cells that resemble myofibroblasts may explain the contractability of the strand. Because the effect of intraocular proliferation can be quantitated by the number of puckers and retinal detachments developing, this model may be useful for the study of therapeutic means to reduce intraocular proliferation.