The blood-retinal barrier permeability to fluorescein in normal subjects and in juvenile diabetics without retinopathy

The blood-retinal barrier permeability to fluorescein was determined in 20 eyes from 17 normal volunteers (mean age 31 years) and in 20 eyes from 19 juvenile diabetics without apparent retinopathy (mean age 35 years - mean duration of diabetes 6 years). The permeability was in normal subjects (1.1 +/- 0.4) X 10(-7) cm/sec (mean +/- 2 X SD) and in juvenile diabetics (1.1 +/- 0.7) X 10(-7) cm/sec (mean +/- 2 X SD). Thus a break-down of the blood-retinal barrier cannot be demonstrated as a very early and general phenomenon in the early course of the diabetic disease. The fluorescein diffusion coefficient in the vitreous body was determined and juvenile diabetics without apparent retinopathy showed a diffusion coefficient of (0.80 +/- 0.25) X 10(-5) cm2/sec (mean +/- 2 X SD), which was the same as in normals where the diffusion coefficient was (0.69 +/- 0.46) X 10(-5) cm2/sec (mean +/- 2 X SD).     

Estimation of the permeability of the blood-retinal barrier in normal individuals

The fluorescein kinetics in the vitreous was simulated with a computer to consider several factors such as permeability of the blood-retinal barrier, outward active transport, plasma fluorescein dynamics, diffusion of fluorescein in the vitreous, and fluorescein leakage from the blood-aqueous barrier. Kinetic vitreous fluorophotometry was performed in normal individuals to estimate the inward and outward permeability of the blood-retinal barrier based on the theory of the simulation model. The results of the simulation studies suggest that the fluorescein concentration in the posterior vitreous after intravenous administration is dependent mainly on the inward permeability and on the plasma concentration and that the outward permeability has little influence on the fluorescein kinetics at the early phase. In the pharmacokinetic analysis of the results of kinetic vitreous fluorophotometry, we obtained average values of 1.8 X 10(-5) cm/min and 5.6 X 10(-4) cm/min for the inward permeability and outward permeability coefficients, respectively. The diffusion coefficient of fluorescein in the vitreous was estimated at 7.9 X 10(-4) cm2/min on the average. The outward permeability of the blood-retinal barrier is approximately 31 times the inward permeability. This suggests that a facilitated process that transports fluorescein outward from the vitreous cavity exists in the blood-retinal barrier of human eyes.     

Quantitative vitreous fluorophotometry applying a mathematical model of the eye

A slit-lamp fluorophotometric method is presented that permits calculation of a blood-retinal barrier permeability to fluorescein (P) and a diffusion coefficient for fluorescein in the vitreous body (D). The calculations are performed by relating the time course of the free--not protein bound--fluorescein concentration in the bloodstream with the fluorescein concentration profile in the vitreous body. The combination is performed automatically on a computer by applying a simplified mathematical model of the eye. P refers to the area of the barrier of the model eye. In a group of six normal persons, the mean P was (1.1 +/- 0.4) X 10(-7) cm/sec (mean +/- SD), while in six diabetic patients with background retinopathy and macular edema the mean P was (7.1 +/- 3.8 ) X 10(-7) cm/sec. The mean D was (7.4 +/- 3.4) X 10(-6) cm2/sec in the normal group and (9.6 +/- 2.0) X 10(-6) cm2/sec in diabetic patients, corresponding as a first approximation to free diffusion in water. Model calculations show that knowing the fluorescein concentration in the bloodstream is considerably significant for the calculation of the permeability, contributing factors up to 50%. For the low-permeation situation, subtraction of the preinjection scan contributes a factor of 50% for both permeability and diffusion coefficient. The exact placement in the vitreous body of the concentration profile, by applying a formalism that transforms slit-lamp movement to intraocular distance, contributes a factor of 20% on the diffusion coefficient. The permeability obtained with the model can be calculated as the ratio between area of vitreous and plasma fluorescein concentration curves within 20%. Active transport of fluorescein across the blood-retinal barrier in the direction of vitreous to blood does not seem to be significant within the first 2 hr after fluorescein injection.     

Blood-retinal barrier permeability to carboxyfluorescein and fluorescein in monkeys

Lipid solubility is a major determinant of permeability across the blood-brain barrier, to which the blood-retinal barrier (BRB) has many similarities. Carboxyfluorescein is a dye with about 1/1000 the lipid solubility of fluorescein, but their molecular sizes and spectral characteristics are similar. We studied the importance of lipid solubility in BRB permeability by comparing the BRB permeabilities to these two dyes. Dye in the vitreous and plasma of four monkeys was measured by fluorophotometry. The estimated inward permeability coefficients (Pin) were 11 +/- 7.4 X 10(-6) cm/min (mean and SD) for carboxyfluorescein and 21 +/- 5.9 X 10(-6) cm/min for fluorescein. The ratio of the means was 1/1.9, far from the expected 1/1000. This finding suggests that the BRB does not function as a continuous lipid membrane and that other factors are more important determinants of permeability for these dyes than lipid solubility.     

Alteration of the blood-retinal barrier and vitreous in sickle cell retinopathy

Nineteen eyes with background sickle cell retinopathy, eleven from patients with SC disease, eight from patients with SS disease, and twelve eyes with proliferative sickle cell retinopathy were examined by direct and indirect ophthalmoscopy, slit-lamp, fluorescein angiography and vitreous fluorophotometry. Calculation of the alteration of the blood-retinal barrier (BRB) and estimation of the diffusion coefficients of fluorescein in the vitreous were performed by fluorophotometry.In background sickle cell retinopathy, the results show a normally functioning BRB in the posterior pole. Abnormally increased fluorescence values to the mid-vitreous (peripheral leakage) were found only in 3 of these 19 eyes, at the two-hour examination (all SC patients). Similarly, mid-vitreous fluorescence values at the two-hour examination were 4.19 ± 1.52 ng/ml in eyes of patients with SC disease, compared with 2.65 ± 0.56 ng/ml in eyes of patients with SS disease. All eyes with background sickle cell retinopathy, except one, showed values for the coefficient of diffusion of fluorescein within normal limits, indicating normal vitreous gel structure.In proliferative retinopathy, the mid-vitreous fluorophotometry readings were abnormally increased, correlating well with the extent of the peripheral angiographic changes (neovascularization). The coefficient of diffusion of fluorescein in the vitreous was generally increased in the eyes with proliferative retinopathy (15.0 ± 8.4 × 10^–4cm²/min) in comparison with a mean value of 5.4 ± 1.4 × 10^–4cm²/min in the eyes wi background sickle cell retinopathy, suggesting an alteration of the vitreous structure eyes with proliferative retinopathy.Fluorophotometry is considered a useful tool to follow patients with sickle cell retinopathy by quantitating peripheral retinal vascular leakage.     

The loss of fluorescein, fluorescein glucuronide and fluorescein isothiocyanate dextran from the vitreous by the anterior and retinal pathways.

The pathways by which fluorescein (F), fluorescein glucuronide (FG) and fluorescein dextran (FD) leave the vitreous body of the rabbit were examined by measuring the concentration distribution of the injected fluorophores in sections of the frozen eyes. The contours of F, as already known, show that it leaves the vitreous predominantly across the retinal surface. Mathematical analysis of the concentration gradient leads to an average outward permeability coefficient of 1.4 x 10(-3) cm min-1 for the retinal layers. The contours of FG and FD show that they leave predominantly by diffusion into the posterior chamber, encountering only a minor barrier at the anterior hyaloid membrane. The anterior contours indicate that there can be no substantial posteriorly directed fluid flow through the vitreous; if it occurs its velocity across the retinal surface must be less than 2 x 10(-5) cm min-1. The contours of FD near the posterior pole of the retina suggest that such a flow may be taking place. Some time after the systemic administration of F, an analysis of the rate of loss of fluorescence from the vitreous body shows that this corresponds to the movement of FG out through the anterior chamber. Its value bears little relationship to the condition of the blood-vitreal barrier.     

Quantitative MR imaging study of intravitreal sustained release of VEGF in rabbits

PURPOSE: To determine whether sustained elevation of vascular endothelial growth factor (VEGF) in the vitreous cavity causes retinal hyperpermeability [blood-retinal barrier (BRB) breakdown] before the development of retinal neovascularization (NV) and to document the kinetics of the integrity of BRB breakdown versus time. METHODS: Poly(L-lactide-co-glycolide)based devices loaded with VEGF were implanted intravitreally in rabbit eyes. Contrast-enhanced magnetic resonance imaging (MRI) methods were used to identify and quantitate the retinal permeability at various time points after implantation. This was done with the newly developed MR tracer AngioMARK (Epix Medical, Boston, MA). After the MRI measurements, fundus photography and fluorescein angiography (FA) also were performed on the same set of animals. RESULTS: At 3 days after implantation, the MR images showed a significant retinal leakage into the vitreous cavity (BRB breakdown) of the VEGF-implanted eyes. To quantitate this leakage, the permeability surface area product (PS) was measured. At 3 days, the mean PS product was 1.25 +/-0.25 x 10(-5) cm3/min. Based on the VEGF in vitro release study, this 3-day BRB breakdown corresponded to a total sustained release of 7.42 +/- 0.54 microg/ml of VEGF. The fundus and FA photographs of these VEGF-implanted eyes taken at 4 days after implantation also showed a considerable level of retinal vascular dilation and tortuosity. By 12 days after implantation, the mean PS product decreased to 5.83 +/- 1.38 x 10(-6) cm3/min. However, the retinal NV was observed only after the second week after implantation. By this time, a total of 10.70 +/- 0.92 microg/ml of VEGF was released in a sustained fashion. Also, after the retinal NV development, retinal detachment also was observed. The control eyes, however, which were implanted with blank devices, remained unchanged and normal during the entire course of this study (PS = 5.57 +/- 0.66 x 10(-7) cm3/min). CONCLUSIONS. The findings indicate that sustained delivery of elevated amounts of VEGF in the vitreous cavity induces a BRB breakdown even earlier than 3 days after implantation. This was achieved after a total sustained release of 7.42 +/- 0.54 microg/ml of VEGF. This retinal leakage regressed by more than half by the time the retinal NV developed. Furthermore, a retinal detachment occurred after this retinal NV. These results are similar to proliferative diabetic retinopathy (PDR). The sustained elevation of VEGF in the vitreous cavity of rabbit eyes is potentially a good model to test VEGF antagonists to treat or prevent PDR in humans. The quantifiable change of BRB breakdown by the contrast-enhanced MRI method is ideal to assess the therapeutic intervention in vivo without killing the animal and may prove to be clinically useful in humans.     

Movement of fluorescein monoglucuronide in the rabbit cornea. Diffusion in the stroma and endothelial permeability

The movement of fluorescein monoglucuronide, a fluorescent metabolite of fluorescein, was studied in the rabbit cornea in vitro and in vivo. A stromal strip was exposed to fluorescein monoglucuronide, and the diffusion rate and the distribution in the stroma were measured every hr for 24 hr. The diffusion coefficient was 0.94 +/- 0.11 (+/- S.D.) X 10(-6) cm2/sec, and the saline/stroma distribution ratio was in a range of 0.67 to 0.69. The concentration of fluorescein monoglucuronide in the anterior chamber and the cornea was measured every hr for 8 hr following intravenous administration. The endothelial permeability was 4.7 +/- 1.0 X 10(-4) cm/min, and the aqueous/cornea distribution ratio was 0.56 +/- 0.05. It appears that the corneal endothelial permeability in the living eye determined hitherto from systemic administration of fluorescein is most likely the permeability to fluorescein monoglucuronide.     

Delivery from episcleral exoplants

PURPOSE: To assess the impact of an episcleral exoplant on transscleral delivery. METHODS: New Zealand White rabbits were given a periocular injection of sodium fluorescein (fluorescein, 376 Da) or an episcleral exoplant loaded with fluorescein. Two types of exoplants were tested: (1) a rigid polyethylene device, impermeable on one side and open to the sclera on the other, that contained compressed pellets of fluorescein and was sutured loosely (apposition group) or tightly to indent the sclera (indentation group) and (2) flexible refillable silicone exoplants also open to the sclera that were secured by suturing, to form a sealed episcleral chamber that was filled with a fluorescein solution. Ocular and plasma fluorophotometry were performed at several time points, and histology was performed to evaluate the effect of exoplants on the periocular tissue. RESULTS: Within 20 minutes of a periocular injection of fluorescein, peak fluorescence was visible in the anterior chamber (AC) and at later time points was displaced toward the retina; at all time points, the highest fluorescence was in the AC. For the polyethylene device indentation group, peak fluorescence was in the retina and posterior vitreous and spread to the AC over time. For the apposition exoplant group, two peaks of fluorescence were seen initially, one in the retina and posterior vitreous and one in the AC. The area under the concentration time curve (AUC +/- SE) for fluorescein concentration was 144.4 +/- 15.1 mug . h/mL for the retinal peak and 43.6 +/- 7.1 mug . h/mL for the posterior vitreous peak after injection of 5 mg of fluorescein into a silicone exoplant, compared with a retinal peak of 3.9 +/- 0.3 and a posterior vitreous peak of 0.99 +/- 0.26 mug . h/mL after periocular injection of 5 mg of fluorescein (P < 0.01 for each). Peak plasma fluorescein levels were significantly reduced in the exoplant group compared with periocular injection. CONCLUSIONS: An episcleral exoplant facilitates diffusion of fluorescein through the sclera resulting in high levels in the retina and posterior vitreous; levels are markedly increased compared with periocular injection of the same amount of fluorescein. It also reduces peak plasma levels indicating reduction of systemic absorption. This procedure provides a new approach that can be combined with sustained-release preparations to optimize delivery of agents to the retina and choroid while minimizing the potential for systemic toxicity.     

Vitreous fluorophotometry in aphakic or pseudophakic eyes with persistent cystoid macular edema

By vitreous fluorophotometry, the degree of disruption of the blood-vitreous barrier was studied in 16 aphakic or pseudophakic eyes with persistent cystoid macular edema (CME) and 11 aphakic or pseudophakic eyes without CME; postoperative periods ranged from 7 to 22 months in both groups. The rate of fluorescein penetration into the vitreous was determined within 30 minutes after intravenous injection. It represented the permeability of the blood-vitreous barrier and averaged 10.28 +/- 4.52 (SD) X 10(-6) min-1 in eyes with CME and 3.05 +/- 1.21 X 10(-6) min-1 in eyes without CME; the difference between the two groups was statistically significant (P less than 0.002, Student t-test). The peak fluorescein concentration in the mid-vitreous (Cv) was determined and the concentration of free fluorescein in the blood serum (Cs) at the corresponding time was also estimated: the Cs/Cv ratio represented the balance between the inward and outward transport of fluorescein across the blood-vitreous barrier. The Cs/Cv ratio was 7.91 +/- 2.94 in eyes with CME and 12.91 +/- 3.68 in eyes without CME: the difference was statistically significant (P less than 0.001). In 6 eyes with the Cs/Cv ration of 5.0-9.2 the condition of CME deteriorated or remained unchanged during the follow-up of 4-8 months, but in 4 eyes with the ratio of 9-14.2 CME showed an improvement during the same period. In 3 eyes with vitreous tug syndrome, anterior vitrectomy improved the ratio from an average of 4.8 to 17.4. It was concluded that a functional disturbance of the blood-vitreous barrier underlies the development of persistent CME.     

Blood-retinal barrier dysfunction at the pigment epithelium induced by blue light.

Exposure to low-intensity white light can induce dysfunction of the blood-retinal barrier (BRB) at the retinal pigment epithelium (RPE). To determine whether the shorter wavelengths white light are responsible for this dysfunction, rabbit retinas were exposed to blue light (400-520 nm) or yellow light (510-740 nm). The permeability of the BRB, a parameter for the integrity of the barrier, was quantified with vitreous fluorophotometry. Morphologically, the barrier at the RPE was visualized on light and electron microscopy using horseradish peroxidase (HRP) as a tracer. Seventeen pigmented rabbits were exposed to blue light and 11 were exposed to yellow light. Vitreous fluorescein leakage increased with the exposure energy according to a power function (correlation coefficient > 0.79). The threshold energy for an increase in BRB permeability was 50 J/cm2 (0.014 W/cm2 for 1 hr) after blue and 1600 J/cm2 after yellow light. HRP tracing demonstrated that after blue light exposure, a significant fluorescein leakage on fluorophotometry corresponded to the presence of HRP in the RPE cells and in the subretinal space. After yellow light exposures of < 3700 J/cm2 and in rabbits with no significant fluorescein leakage, the HRP was limited to the choroidal capillaries and Bruch's membrane. These results demonstrate that the blue component of white light causes dysfunction of the BRB at the RPE 30 times more effectively than the longer wavelength fraction of white light. As a result, a blue light blocking filter should be used in ocular surgery on humans when an operating microscope is being used (light power 0.1-0.9 W/cm2).     

Pharmacokinetic interpretation of vitreous fluorophotometry

Vitreous fluorophotometry is producing an increasing amount of clinical and experimental data. In order to interpret these data and obtain quantitative values for the permeability of the blood ocular barriers, there is a need to understand better the basic phenomena governing the transport of fluorescein. We present here a refined mathematical model that we use to interpret a large body of clinical data yielding values for the inward (6.9 x 10(-6) cm/min) and outward (210 x 10(-6) cm/min) posterior permeability coefficients, the effective diffusion coefficient in the vitreous (8 x 10(-4) cm2/min), and the plasma fluorescein decay constants (1.17, 0.34, and 0.044 per hour). Moreover, we utilize the model to make predictions related to kinetic vitreous fluorophotometry and to the reliability of the procedure to calculate the permeability coefficients.     

Blood-retina barrier permeability in diabetes during acute ACE-inhibition.

We assessed the acute effect of ACE-inhibition (captopril) on blood-retina barrier (BRB) permeability in 10 hypertensive insulin-dependent diabetic patients with background retinopathy in a double-masked placebo controlled cross-over study. All patients underwent ophthalmological examination, fundus photography, fluorescein angiography, vitreous fluorometry, and continuous blood pressure recording within 3 h of the drug/placebo administration. The decrease in mean arterial blood pressure, from placebo treatment 149/92 +/- 17/7 to captopril treatment 132/83 +/- 14/7 mmHg (mean +/- SD), P less than 0.01 was not accompanied by a significant decrease in BRB permeability, which was 2.51 (1.24-9.15) with placebo and 3.02 (1.25-13.93).10(-7) cm/s during captopril treatment (geometric mean and-range), NS. Our study suggests that abnormal leakage through the BRB in hypertensive insulin-dependent diabetic patients with background retinopathy is caused predominantly by structural changes in the retinal vessels whereas hydrostatic forces play a minor role.     

Quantitative control of the function of the corneal endothelium by fluorophotometry in the anterior eye segment

Whereas specular microscopy yields mostly qualitative information, fluorophotometry furnishes quantitative data of endothelial cell function. Determination of fluorescein permeability of the endothelial cell layer reflects the function of the endothelial barrier. Following topical application of fluorescein, the time-dependent change in the fluorescein concentration in corneal stroma and aqueous yields the transfer coefficient (Kc) of the corneal endothelium. With the Fluorotron Master, Kc was 3.77 +/- 0.57 X 10(-3)/min. for normal eyes; patients with cornea guttata or Fuchs's dystrophy had a significantly higher transfer coefficient (Kc = 7.9 +/- 2.88 X 10(-3)/min.). Normal Kc values were found 6-63 months (average 33.5 months) after phacoemulsification with implantation of a posterior chamber lens.     

Effect of acetazolamide on outward permeability of blood-retina barrier using differential vitreous flyorophotometry.

PURPOSE: To measure fluorescein (F) and fluorescein monoglucuronide (FG) concentrations in the vitreous and evaluate the effect of acetazolamide (AZM) on the outward permeability of the blood-retina barrier (BRB) using differential vitreous fluorophotometry (DVF). METHODS: DVF was performed 180 minutes after intravenous injection of AZM (5 mg/kg) and 50 mg of sodium fluorescein in six rabbits (AZM group). DVF also was performed in six rabbits injected intravenously with only 50 mg of sodium fluorescein (control group). The F/FG ratio was calculated based on the concentrations of F and FG obtained by DVF. DVF also was performed 180 minutes after 50 mg of intravenous injection of sodium fluorescein in five rabbits given probenecid (150 mg/kg) intraperitoneally (probenecid group). RESULTS: The average F/FG ratio was 0.36 +/- 0.17 (range, 0.22-0.66) in the AZM group, which was significantly smaller than the control value of 0.74 +/- 0.22 (range, 0.50-1.60). The average F/FG ratio at 180 minutes after injection was 1.51 +/- 0.46 (range, 0.94-2.00) in the probenecid group, which was significant higher (p < 0.05) than that of the AZM or control group. CONCLUSIONS: This study showed that the F/FG ratio might be a good indicator of the estimated outward permeability of the BRB using DVF and that AZM may accelerate the outward active transport function of the BRB.     

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.     

Posterior vitreous fluorophotometry in diabetic patients with minimal or no retinopathy

Vitreous fluorophotometry (VFP) using the Fluorotron Master was performed in 25 insulin-dependent diabetic patients with either minimal or no retinopathy and 22 controls. At 30 minutes, baseline corrected vitreous fluorescence values 3 mm from the retina were significantly greater in diabetic patients with minimal retinopathy than in either controls or patients with no retinopathy but were similar in patients with no retinopathy and controls. At 1 hour, vitreous fluorescence values and fluorescein permeability indexes were similar in all three groups. The absence of increased posterior vitreous fluorescein leakage in the diabetic patients with no retinopathy suggests that the alteration in permeability of the blood retinal barrier, as assessed by current VFP measures, does not precede the development of clinical diabetic retinopathy. However, the significantly increased 30-minute 3-mm fluorescence values suggest that posterior fluorescein leakage is increased in minimal background diabetic retinopathy.     

Movement of fluorescein and fluorescein glucuronide across the isolated rabbit iris-ciliary body

Movement of fluorescein and fluorescein glucuronide, a fluorescent metabolite of fluorescein, across the isolated iris-ciliary body of the albino rabbit was determined under short-circuit conditions using a modified Ussing's chamber. The permeabilities of this tissue to these dyes were calculated. The outward permeability (from the aqueous to the stromal side) of the iris-ciliary body preparation averaged 6.63 +/- 0.86 for fluorescein and 1.51 +/- 0.47 X 10(-6) cm/sec for fluorescein glucuronide, and the inward permeability (from the stromal to the aqueous side) was 1.68 +/- 0.41 for fluorescein and 1.37 +/- 0.77 X 10(-6) cm/sec for fluorescein glucuronide, respectively. Application of probenecid or ouabain decreased the outward permeability of fluorescein, but it had no significant effect on the fluorescein glucuronide movement. Application of 10(-5) M 2,4-dinitrophenol showed no significant effect on the fluorescein or fluorescein glucuronide movement, but application of 5 X 10(-4) M 2,4-dinitrophenol decreased the outward fluorescein transfer, which was also markedly suppressed by incubation at 0 degrees C. It is possible that an active transport mechanism is involved in the outward fluorescein movement across the iris-ciliary body, while the inward movement of fluorescein and also the fluorescein glucuronide movement across this tissue is mainly by passive diffusion.     

Transport of fluorescein in the rabbit eye after treatment with sodium iodate

The outward active transport and the inward permeability of the blood-retinal barrier were studied in the rabbit eye after i.v. administration of sodium iodate. The active transport was evaluated from the half-time of disappearance of the vitreous fluorescein following intravitreal administration, and the inward permeability was evaluated from the vitreous concentration of fluorescein monoglucuronide after i.v. administration. The half-time of the vitreous fluorescein was 3.5 +/- 0.3 (mean +/- S.D.) hr, and 3.9 +/- 0.2 hr before and within 6 hr after iodate administration, respectively. After 24 hr, the half-time was 11.7 +/- 1.7 hr, similar to that of fluorescein monoglucuronide, 12.0 +/- 2.7 hr. The vitreous and the anterior chamber concentration of fluorescein monoglucuronide was measured at 1 hr after the i.v. dye injection. The vitreous concentration in the rabbits given iodate 3 hr before the dye injection was significantly greater than in the normal eyes, while the anterior chamber concentration was not different. Since fluorescein is rapidly metabolized to fluorescein monoglucuronide, differences in parameters determined using systemic fluorescein under two treatments or in disease states may be the result of alteration of the dynamics of fluorescein, fluorescein monoglucuronide, or both.     

Diabetic macular edema: passive and active transport of fluorescein through the blood-retina barrier

PURPOSE: To investigate the passive bidirectional and active outward transport of fluorescein through the blood-retina barrier (BRB) in diabetic patients with clinically significant macular edema and in healthy controls. METHODS: The passive and active transport of fluorescein through the BRB was quantitated by vitreous fluorometry. A previously developed method was used to model passive transport. A new simulation model was developed and evaluated for estimation of active transport. The study included 10 eyes of 5 healthy controls and 31 eyes of 20 diabetic patients with clinically significant diabetic macular edema (CSME) in at least one eye, totalling 25 eyes with CSME. RESULTS: Passive permeability of fluorescein was increased by a factor of 12 in eyes with edema compared to healthy controls (edema, 23.7 nm/sec; healthy subjects, 1.9 nm/sec, P < 0.01), whereas the active transport was doubled (edema, 84.1 nm/sec; healthy subjects, 43.5 nm/sec, P < 0.01). Unlike active transport, passive permeability was related to the degree of retinopathy, in that eyes with severe non-proliferative diabetic retinopathy had a passive permeability that was significantly increased compared to moderate retinopathy (32.1 nm/sec and 14.6 nm/sec, respectively, P: < 0.05). The passive movement quantitated with vitreous fluorometry was larger for diffuse and mixed leakage compared to focal (P = 0.07). CONCLUSIONS: Insofar as the movement of fluorescein can be taken as a probe for the movement of electrolytes and water, the pathogenesis of diabetic macular edema seems to involve a disruption of the BRB, presumably its inner component. The active resorptive functions of the blood-retina barrier appear to be compensatorily increased to counteract edema formation, although the increase is too small to prevent edema in the face of severe leakage through the blood-retina barrier.