Manganese-Enhanced MRI of the Brain in Healthy Volunteers.

BACKGROUND AND PURPOSE: The manganese ion is used as an intracellular MR imaging contrast agent to study neuronal function in animal models, but it remains unclear whether manganese-enhanced MR imaging can be similarly useful in humans. Using mangafodipir (Teslascan, a chelated manganese-based contrast agent that is FDA-approved), we evaluated the dynamics of manganese enhancement of the brain and glandular structures in the rostral head and neck in healthy volunteers. MATERIALS AND METHODS: We administered mangafodipir intravenously at a rate of 1 mL/minute for a total dose of 5 µmol/kg body weight. Nine healthy adult volunteers (6 men/3 women; median age, 43 years) completed baseline history and physical examination, 3T MR imaging, and blood work. MR imaging also followed mangafodipir administration at various time points from immediate to 7 days, with delayed scans at 1-3 months. RESULTS: The choroid plexus and anterior pituitary gland enhanced within 10 minutes of infusion, with enhancement persisting up to 7 and 30 days, respectively. Exocrine (parotid, submandibular, sublingual, and lacrimal) glands also enhanced avidly as early as 1 hour postadministration, generally resolving by 1 month; 3 volunteers had residual exocrine gland enhancement, which resolved by 2 months in 1 and by 3 months in the other 2. Mangafodipir did not affect clinical parameters, laboratory values, or T1-weighted signal in the basal ganglia. CONCLUSIONS: Manganese ions released from mangafodipir successfully enable noninvasive visualization of intra- and extracranial structures that lie outside the blood-brain barrier without adverse clinical effects, setting the stage for future neuroradiologic investigation in disease.

Validation of structural and functional lesions of diabetic retinopathy in mice.

Diabetic retinopathy is a serious long-term complication of diabetes mellitus. There is considerable interest in using mouse models, which can be genetically modified, to understand how retinopathy develops and can be inhibited. Not all retinal lesions that develop in diabetic patients have been reproduced in diabetic mice; conversely, not all abnormalities found in diabetic mice have been studied or identified in diabetic patients. Thus, it is important to recognize which structural and functional abnormalities that develop in diabetic mice have been validated against the lesions that characteristically develop in diabetic patients. Those lesions that have been observed to develop in the mouse models to date are predominantly characteristic of the early stages of retinopathy. Identification of new therapeutic ways to inhibit these early lesions is expected to help inhibit progression to more advanced and clinically important stages of retinopathy.

Critical role of inducible nitric oxide synthase in degeneration of retinal capillaries in mice with streptozotocin-induced diabetes.

AIMS/HYPOTHESIS: Diabetes results in the upregulation of the production of several components of the inflammatory response in the retina, including inducible nitric oxide synthase (iNOS). The aim of this study was to investigate the role of iNOS in the pathogenesis of the early stages of diabetic retinopathy using iNOS-deficient mice (iNos (-/-)). MATERIALS AND METHODS: iNos (-/-) mice and wild-type (WT; C57BL/6J) mice were made diabetic with streptozotocin or kept as non-diabetic controls. Mice were killed at different time points after the induction of diabetes for assessment of vascular histopathology, cell loss in the ganglion cell layer (GCL), retinal thickness, and biochemical and physiological abnormalities. RESULTS: The concentrations of nitric oxide, nitration of proteins, poly(ADP-ribose) (PAR)-modified proteins, endothelial nitric oxide synthase, prostaglandin E(2), superoxide and leucostasis were significantly (p < 0.05) increased in retinas of WT mice diabetic for 2 months compared with non-diabetic WT mice. All of these abnormalities except PAR-modified proteins in retinas were inhibited (p < 0.05) in diabetic iNos (-/-) mice. The number of acellular capillaries and pericyte ghosts was significantly increased in retinas from WT mice diabetic for 9 months compared with non-diabetic WT controls, these increases being significantly inhibited in diabetic iNos (-/-) mice (p < 0.05 for all). Retinas from WT diabetic mice were significantly thinner than those from their non-diabetic controls, whereas diabetic iNos (-/-) mice were protected from this abnormality. We found no evidence of cell loss in the GCL of diabetic WT or iNos (-/-) mice. Deletion of iNos had no beneficial effect on diabetes-induced abnormalities on the electroretinogram. CONCLUSIONS/INTERPRETATION: We demonstrate that the inflammatory enzyme iNOS plays an important role in the pathogenesis of vascular lesions characteristic of the early stages of diabetic retinopathy in mice.

Correction of early subnormal superior hemiretinal DeltaPO(2) predicts therapeutic efficacy in experimental diabetic retinopathy.

PURPOSE: To test the hypothesis that regional retinal oxygenation responses to a hyperoxic inhalation challenge are associated with reported retinopathy outcomes after different therapies in rat models of diabetic retinopathy. METHODS: Six groups of rats were maintained for 3 months: controls (n = 8), untreated diabetic (n = 8), aminoguanidine (AMG)-treated diabetic (2.5 g/kg of diet; n = 6), untreated galactosemic (n = 7), AMG-treated galactosemic (n = 10), and WAY-509-treated (25 mg/kg body weight per day) galactosemic (n = 7). After 3 months, the change in oxygen tension was measured noninvasively from the superior to the inferior ora serrata, using a novel functional magnetic resonance imaging (fMRI) technique and a carbogen (a gas mixture of 5% carbon dioxide and 95% oxygen that has been used clinically, instead of 100% oxygen, to minimize the vasoconstrictive effects of pure O(2) on retinal blood flow and oxygenation) inhalation challenge. Retinal morphometric measurements were also obtained. RESULTS: Retinal lesions (acellular capillaries and pericyte ghosts) were not significantly (P > 0.05) present at 3 months in any experimental groups compared with the control group. Superior but not inferior hemiretinal change in partial pressure of oxygen (DeltaPO(2)) became significantly subnormal (P < 0.05) at 3 months of diabetes or galactosemia. Aminoguanidine, which has been found to inhibit the development of retinopathy in diabetic but not galactosemic rats, inhibited the development of a subnormal DeltaPO(2) in diabetes but not in galactosemia. WAY-509, which has been reported to inhibit retinopathy in galactosemic rats, inhibited the DeltaPO(2) defect in galactosemic rats. CONCLUSIONS: An early subnormal superior hemiretinal DeltaPO(2) after treatment appears to be a good predictor of the risk of development of retinopathy, as well as for assessing therapeutic efficacy in experimental diabetic retinopathy.

Variable supplemental oxygen during recovery does not reduce retinal neovascular severity in experimental ROP.

PURPOSE: To test the hypothesis that variable supplemental oxygen (VSO) during recovery is not as effective as constant supplemental oxygen treatment at reducing retinal neovascular severity in the newborn rat model of retinopathy of prematurity (ROP). METHODS: The ROP model consists of raising newborn rats in variable oxygen for 14 days and then allowing them to recover during the next 6 days (until day 20). During this recovery period, two VSO protocols were examined: 1) 24 hr cycled (28% for 24 hr and 21% for 24 hr) as a control, and 2) interrupted (28% for 23.75 hr and 21% for 0.25 hr). On day 20, retinas from both eyes of each animal were dissected, stained with ADPase, and analyzed for neovascular (NV) incidence, severity, and peripheral avascularity. RESULTS: No significant differences (P > 0.05) in NV incidence and severity were found between the 24 hr cycled (control) (95% (38/40), median 5 clockhrs, range 0.5-9, n = 38) and interrupted VSO groups (89% (40/45), 3.5 clockhrs, 0.5-10.5, n = 40). However, retinal NV severity from both VSO groups were significantly greater (P < 0.05) than that previously reported for constant supplemental oxygen treatment. Percent peripheral avascularities were not significantly different (P > 0.05) between the two VSO groups. CONCLUSIONS: These results support our hypothesis since both VSO conditions were significantly less effective at reducing retinal NV than constant supplemental oxygen during recovery. The possible application of these results to STOP-ROP trial is discussed.

Measuring the human retinal oxygenation response to a hyperoxic challenge using MRI: eliminating blinking artifacts and demonstrating proof of concept.

The retinal oxygenation response to a hyperoxic challenge measured using MRI appears to be an early and accurate marker of retinopathy risk in experimental models, with promising clinical potential. However, the application of this technique in humans is limited by blinking artifacts that can confound detection of subtle signal intensity changes. We asked subjects to refrain from blinking during a 12-s fast low-angle shot (FLASH) image, and to blink if needed during the following 3-s rest period. This no-blink blink cycle was repeated sequentially 20 times during either room-air or 100% oxygen breathing. Significant change (P < 0.05) was detected for the first time from the resultant blinking-artifact-free images in the preretinal vitreous oxygen tension (upper limit of about 13 mm Hg (1.8 KPa, N = 3)) following a 10-min hyperoxic inhalation challenge. These results provide the proof-of-concept data needed for future MRI evaluation of the retinal oxygenation response and human retinopathy, such as diabetic retinopathy.

MR studies of retinal oxygenation.

In this paper, we summarize the development and application of two novel magnetic resonance based measurements of retinal oxygenation in experimental models of retinopathy, including diabetic retinopathy and retinopathy of prematurity. We use 19F-NMR and a small (microl) perfluorocarbon drop positioned in the preretinal vitreous space to make PO2 measurement of the inner retina. In addition, we use magnetic resonance imaging (MRI) to accurately and non-invasively measure the change in the preretinal PO2 (DeltaPO2) following the shift from breathing room air to a hyperoxic inhalation challenge. The advantages and disadvantages of each method are discussed. New applications of these techniques in the newborn rat and adult mouse are presented. We expect such studies to motivate future MRI oxygenation studies of human retinopathy, including diabetic retinopathy and retinopathy of prematurity.

Preretinal neovascularization associated with acetazolamide-induced systemic acidosis in the neonatal rat.

PURPOSE: NH4Cl gavage in the neonatal rat produces a metabolic acidosis-induced retinopathy which serves as a model for retinopathy of prematurity (ROP). Acetazolamide induces a metabolic acidosis via an alternative biochemical mechanism (bicarbonate loss versus hydrogen ion load). In the present study, the following hypothesis was tested: acetazolamide-induced acidosis is associated with preretinal neovascularization in the neonatal rat. METHODS: All studies used newborn Sprague-Dawley rats raised in expanded litters of 25. Arterial blood pH was measured to determine the level of acidosis induced by intraperitoneal (IP) acetazolamide (50 or 200 mg/kg) or saline. In a separate retinopathy study, newborn rats (n = 75) were randomized to either IP acetazolamide, 50 mg/kg (low-dose), or IP saline twice daily from days 2 to 7. After 5 days of recovery, retinal vasculature was assessed using ADPase staining and light microscopy. The presence and severity (clock hours) of neovascularization were assessed by three masked observers. In an additional retinopathy study, newborn rats (n = 100) were randomized to either IP acetazolamide, 200 mg/kg (high-dose), or IP saline twice daily from days 2 to 7. After 5 days of recovery, the retinas were similarly analyzed. RESULTS: Neovascularization occurred in 59% of rats receiving high-dose acetazolamide (200 mg/kg). High-dose acetazolamide produced a severe acidosis (pH 7.13 +/- 0.06) during drug delivery. Low-dose acetazolamide (50 mg/kg) produced a pH (7.22 +/- 0.07) that was intermediate between high-dose (200 mg/kg) acetazolamide (P < 0.001) and saline controls (7.42 +/- 0.06, P < 0.001); however, neither low-dose acetazolamide nor saline induced preretinal neovascularization. CONCLUSIONS: Acidosis induced by high-dose acetazolamide, independent of hyperoxemia or hypoxemia, is associated with preretinal neovascularization in the neonatal rat. Induction of neovascularization appears to depend on a critical threshold of acidosis severity. This study further supports a proposed independent role for acidosis in the pathogenesis of ROP.

The effect of partial vitrectomy on blood-ocular barrier function in the rabbit.

PURPOSE: To compare ocular vascular permeability in the rabbit after vitrectomy as assessed by contrast-enhanced magnetic imaging (CE-MRI) and measurements of aqueous and vitreous humor protein concentration. METHODS: Partial vitrectomies were performed, irrigating with BSS or BSS PLUS. Post-operative vascular leakage was determined by CE-MRI following intravenous administration of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA). Aqueous and vitreous protein concentrations were quantified by standard biochemical assay. ERG evaluations were performed on postoperative days 1, 3, and 7. RESULTS: Using BSS as irrigant, breakdown of the inner blood-retinal barrier (BRB) occurred in 4/7 eyes on post-operative day 1. The rate of Gd-DTPA leakage was significantly greater on postoperative day 1 than that in unoperated, control eyes, but declined approximately 50% by day 3. At both time points, outer BRB breakdown was restricted to the sclerotomy wounds. No BRB leakage was detectable in control eyes. Blood-aqueous barrier (BAB) leakage was bilateral on day 1. Significantly greater Gd-DTPA leakage occurred in the operated eye than in the nonsurgical contralateral eye. On day 3, approximately 40% bilateral reduction in leakage indicated resolution of BAB leakage. Notably, Gd-DTPA leakage of the BAB and BRB was significantly reduced in the BSS PLUS treated group. In contrast to MRI assessments, protein concentrations of the aqueous and vitreous in the surgical eye showed no detectable differences between BSS and BSS PLUS. Concurrent with the transient loss of ocular barrier function, ERG responses also declined. However, by day 7 greater than 90% recovery was noted in BSS PLUS treated animals but not in the BSS treatment group. CONCLUSIONS: CE-MRI is capable of detecting subtle changes in vascular permeability following ocular surgery. Advantages of using BSS PLUS compared to BSS as the irrigating solution can be detected using this technique. BSS PLUS's protection of barrier function is consistent with a rapid recovery in retinal function not observed in BSS treated eyes.

Significant reduction of the panretinal oxygenation response after 28% supplemental oxygen recovery in experimental ROP.

PURPOSE: To test the hypothesis that after supplemental oxygen recovery (SOR) in the newborn rat model of retinopathy of prematurity (ROP) the preretinal neovascular (NV) incidence and severity are decreased and the panretinal oxygenation ability is improved. METHODS: Newborn rats were first raised in either room air (controls) or variable oxygen (50%/10%) for 14 days. The experimental rats were recovered during the next 6 days (until day 20) in either room air (21% O2) or supplemental oxygen (28%). All groups were then exposed to room air for an additional 6 days (until day 26). On day 20, magnetic resonance imaging (MRI) was used to determine the panretinal oxygenation response (deltaPO2, mm Hg) to a carbogen (95% O2/5% CO2) inhalation challenge. On days 20 and 26, the retinas from a different subset of control, room air-recovered, or SOR-recovered animals were analyzed using ADPase stained or fluorescein-labeled dextran infused retinal flatmounts. RESULTS: On day 20, the panretinal deltaPO2 of the room air-recovered group (125 +/- 5 mm Hg, mean +/- SEM, n = 12) was significantly (P < 0.05) lower than that of the control group (179 +/- 6 mm Hg, n = 11). The panretinal deltaPO2 value for the SOR group (87 +/- 5 mm Hg, n = 7) was significantly (P < 0.05) lower than both the room air-recovered group and the control group. The NV incidence and severity were significantly reduced (P < 0.05) in the SOR animals compared with the room air-recovered animals. In contrast, on day 26 (after 6 days in room air), the NV incidence was statistically (P < 0.05) greater in the animals that had been exposed to SOR compared with room air-recovered animals. CONCLUSIONS: After 28% SOR, the expected decrease in NV incidence and severity occurred but with an unexpected decrease in panretinal oxygenation ability. The present data strongly support an association between subnormal panretinal oxygenation ability and increased NV risk in the newborn rat ROP model. MRI appears to be a powerful new approach for quantitatively and noninvasively measuring retinal oxygenation and may be applicable to study other ischemic or ischemia-related retinopathies in addition to ROP, such as diabetic retinopathy, sickle cell retinopathy, macular degeneration, and glaucoma.

Subnormal retinal oxygenation response precedes diabetic-like retinopathy.

PURPOSE: Determining which patients are at risk for the development of diabetic retinopathy is expected to greatly improve existing prevention and treatment options. In this study, using an animal model of diabetic retinopathy, the hypothesis was tested that magnetic resonance imaging (MRI) and a carbogen inhalation challenge provides important diagnostic information regarding the risk of developing diabetic retinopathy. METHODS: MRI was used to measure noninvasively the change in oxygen tension along the entire inner retina (i.e., from superior ora serrata to inferior ora serrata) during a carbogen (95% O2/5% CO2) inhalation challenge (IOVS 1996;37:2089). Two animal groups were examined by this MRI method at two time points: (1) rats fed either normal rat chow (n = 20) or a 50% galactose diet (n = 20) for 3.5 months (i.e., before the appearance of extensive retinal lesions) or (2) rats fed either normal rat chow (n = 3) for 15 months or a 30% galactose diet (n = 4) for 15 to 18 months (i.e., when lesions are present). Retinal biochemical and morphometric measurements were also obtained. RESULTS: After 3.5 months of galactosemia, before the appearance of extensive retinal morphologic lesions, a significant (P < 0.05) reduction in the panretinal oxygenation response was observed in the galactosemic group compared with its age-matched control. These galactose-fed animals also displayed a significantly (P < 0.05) larger oxygenation response in the inferior hemiretina than in the superior hemiretina. After 15 to 18 months of galactosemia, during the period when lesions are present, the panretinal oxygenation response remained significantly (P < 0.05) lower in the galactose-fed animals than in their age-matched controls. In contrast to the 3.5-month results, the oxygenation response in galactosemic animals at 15 to 18 months was significantly (P < 0.05) larger in the superior than in the inferior hemiretina. Hemiretinal oxygenation responses were not different in normal controls at either duration. CONCLUSIONS: MRI measurement of the retinal oxygenation response to a carbogen challenge appears to be a powerful new and noninvasive approach that may be useful for assessing aspects of pathophysiology underlying the development of diabetic retinopathy in galactosemic rats. These results support our working hypothesis and suggest that further research into the diagnostic potential of this MRI approach for predicting the development of diabetic retinopathy is warranted.

Angiographic criteria reliably predict when carotid endarterectomy can be safely performed without a shunt.

BACKGROUND: Selective shunting during carotid endarterectomy is widely performed, but the optimal approach for predicting when a shunt is unnecessary remains uncertain. We evaluated the ability of preoperative cerebral angiography to predict when carotid endarterectomy could be safely performed without a shunt. STUDY DESIGN: Eighty-seven patients undergoing carotid endarterectomy between August 1991 and December 1997 had preoperative cerebral angiograms. The angiograms were evaluated for the presence of collateral flow from the contralateral carotid through the anterior communicating artery and from the posterior circulation through the posterior communicating artery. Patients then underwent endarterectomy and were selectively shunted based on somatosensory evoked potential changes. Internal carotid artery stump pressure was routinely measured in all patients. RESULTS: Nine patients (10%) had a shunt placed based on somatosensory evoked potential changes and none of the 87 patients had a perioperative (30 days) stroke. Angiography revealed that 36 patients (41%) had no cross-filling from the contralateral carotid through the anterior communicating artery. Nine of these patients (25%) required a shunt; none of the 51 patients with adequate cross-filling (p < 0.001) did. Furthermore, 94% of the patients without cross-filling but with a patent ipsilateral posterior communicating artery did not require a shunt using somatosensory evoked potential changes as the standard for shunt insertion. Stump pressure measurements (> or = 25 mmHg) or (> or = 50 mmHg) did not reliably exclude the need for a shunt. Only 2 of 15 patients with contralateral carotid occlusion and 1 of 16 patients with a prior ipsilateral stroke required shunts. CONCLUSIONS: In the presence of cross-filling from the contralateral carotid artery, shunt insertion was uniformly unnecessary. In addition, routine shunting of patients with previous ipsilateral strokes or contralateral carotid occlusion was not always necessary. Stump pressures were less sensitive than angiographic criteria in determining when a shunt was unnecessary. Evaluation of cross-filling from the contralateral carotid artery on preoperative angiography can predict with certainty which patients will not require a shunt.

Electron microscopic evidence for the mechanism of blood-retinal barrier breakdown in diabetic rabbits: comparison with magnetic resonance imaging.

Diabetes leads to a breakdown of the blood-retinal barrier (BRB), which can be demonstrated in experimental models by immunocytochemistry and magnetic resonance imaging (MRI). The present study utilizes these methods to investigate the mechanism of BRB breakdown in diabetic rabbits, a model ideally suited to both procedures. Rabbits were treated with alloxan and examined 2 months, 1 year, and 1.5 years after the development of diabetes to assess BRB breakdown using MRI and immunocytochemical staining for endogenous albumin. Using MRI, an increased incidence of retinal vascular leakage is first evident at 1 year of diabetes. Electron microscopic immunolocalization of albumin suggests that BRB compromise is principally mediated by transendothelial transport of serum proteins in endocytic vesicle-like structures of approximately 0.4-1 micron diameter. Some additional retinal vascular leakage is occasionally demonstrated through the interendothelial cell tight junctions, but only when adjacent vascular endothelial cells show degenerative changes. The similarity of these findings to those previously reported for diabetic humans and rats supports the use of the diabetic rabbit as a model for studying BRB dysfunction. MRI and electron microscopic (EM) immunocytochemistry are complementary methods for evaluating BRB dysfunction. MRI can provide an overall picture of the entire eye without sacrificing the animal. EM immunocytochemistry can provide a more detailed picture of a limited area of interest to gain insight into the mechanisms of extravasation. Together, both methods provide a more complete understanding of BRB breakdown in diabetic rabbits.

Abnormal panretinal response pattern to carbogen inhalation in experimental retinopathy of prematurity.

PURPOSE: Present technologies are not able to determine which retinas are at risk for the development of neovascularization in retinopathy of prematurity (ROP). In this study, the authors evaluated whether a novel magnetic resonance imaging (MRI) method could be used to identify differences between control retinas and those that will develop neovascularization in the newborn rat model of retinopathy of prematurity (ROP). METHODS: MRI and a 2-minute carbogen (95% O2/5% CO2) inhalation challenge (see ref. 11) were used to measure noninvasively the change in the posterior vitreous oxygen tension in specific locations across the full extent of the retina in day-12 rats raised in either room air (control, n = 7) or variable oxygen conditions (experimental ROP, n = 7). The experimental ROP animals were examined 2 days before the onset of neovascularization. RESULTS: In the ROP group, the response to carbogen was lower (P < 0.05) at every distance from the optic nerve than in the control group. Within the ROP group, the vascular midperipheral retinal reaction to carbogen, 1 to 2 mm from the optic nerve, was as low as that from the avascular periphery, 2 to 3 mm from the optic nerve. Although the vascular central retinal response to carbogen, 0 to 1 mm from the optic nerve, was greater than either the vascular midperipheral retina or the avascular periphery in the ROP group, theoretically this difference could be caused by oxygen diffusing from the hyaloidal circulation. CONCLUSIONS: Carbogen-challenge MRI seems to be a useful tool for assessing the risk of retinal neovascularization in the newborn rat ROP model. This MRI method has potential clinical applicability, for example, because effective laser therapy with retinal sparing may be possible if focal photocoagulation, guided by an MRI map, is performed.

The vitreous protein concentration is increased prior to neovascularization in experimental ROP.

PURPOSE: To test the hypothesis that the vitreous protein content is altered prior to the development of neovascularization in an experimental model of retinopathy of prematurity (ROP). METHODS: Newborn rats underwent either a variable oxygen exposure or room air exposure following birth. On day 13 or 14, two days prior to neovascularization, the steady state vitreous and plasma total protein levels in room-air controls or variable-oxygen-exposed newborn rats were determined; similar measurements were also made for control adult rats. RESULTS: There was a significant difference (P < 0.05, 2-tailed t-test) in the vitreous protein concentration between the age-matched control and experimental newborn rats. The protein level in the ROP rat plasma was not significantly (P > 0.05) different from that in the age-matched control animals. The vitreous-to-plasma-protein ratios of both the control and ROP newborn animals were significantly greater (P < 0.05) than that in the adults. CONCLUSIONS: The results of this study demonstrate, for the first time, increased vitreous protein levels prior to the development of neovascularization in the newborn rat model of ROP. In addition, developmental changes in vitreous protein levels were identified. The role of developmental and pathologic alterations in the blood-ocular barriers in this study is discussed.

Impaired hyaloidal circulation function and uncoordinated ocular growth patterns in experimental retinopathy of prematurity.

PURPOSE: To test the hypotheses that, in the newborn rat model of retinopathy of prematurity (ROP), the hyaloidal circulation is functionally impaired and its development is not well coordinated with that of other ocular structures. METHODS: The functional response of the hyaloidal circulation to a carbogen inhalation challenge was noninvasively evaluated using magnetic resonance imaging (MRI) in day 12 rats raised under either variable oxygen conditions (experimental ROP, n = 8) or room air (control, n = 8). A similar MRI examination was performed in separate experiments using either day 18 newborn control rats (n = 3) or adult rats (n = 9). For each experiment, the hyaloidal circulation perfusion response to carbogen, the functional spatial extent of the hyaloidal circulation in vitreous, and the volumes of vitreous and lens were estimated from MRI enhancement maps. RESULTS: The hyaloidal perfusion response to carbogen breathing in the newborn rats decreased as follows: control day 12 > experimental day 12 > control day 18; no measurable hyaloidal function was found in the adult rat. Regression analysis indicated a relatively poorer superior-inferior correlation in the temporal response to carbogen inhalation for the experimental animals than in the control newborn rats. The vitreous volume decreased in control rats as expected (adult rat > day 18 > day 12). Good agreement was found between the MRI-determined adult rat vitreous volume (56 +/- 2 microliters) and that of previous reports. Functional hyaloidal volumes during carbogen breathing were not significantly different (P > 0.05) between day 18, day 12 control, and experimental newborn rats. The ratio of this functional hyaloidal circulation extent volume to vitreous volume was significantly different (P < 0.05) between these groups. Covariance analysis revealed a relatively less coordinated development between the functional hyaloidal volume and the vitreous volume in experimental animals than in age-matched control animals, whereas there was coordinated evolution of the hyaloidal circulation and the lens in all the animals. CONCLUSIONS: Carbogen-enhanced MRI appears to be a powerful new and noninvasive approach for assessing the functionality of the hyaloidal circulation (that is, its ability to respond to a carbogen challenge) and quantitatively comparing the functional hyaloidal extent to other ocular volumes in the same eye during development and during the disease process. Evidence is presented here for the first time that supports the authors' hypotheses that the function of the hyaloidal circulation in experimental ROP is impaired and that the growth of ocular components are less coordinated.

A comparative study of the effects of argon and diode laser photocoagulation on retinal oxygenation.

BACKGROUND: The purpose of this study was to compare the effects of diode and argon laser photocoagulation (DLP and ALP) on the preretinal oxygen tension (PO2): (1) directly over photocoagulated retina and (2) in between laser lesions. METHODS: DLP or ALP was applied to avascular rabbit retina to produce grade II lesions. On the day of the oxygen measurement, a droplet of perfluorotributylamine was placed into the preretinal vitreous space over the lasered area of retina and the steady-state PO2 was measured in normoxic animals using 19F magnetic resonance spectroscopy. To determine the PO2 directly over laser lesions, small (5-microliter) droplets were placed over large (approximately 4 mm x 5 mm), confluent areas of treatment (burn area approximately 95% of the treated retinal surface area). To determine the PO2 in between laser lesions, a large (10-microliter) droplet was placed over a field of scatter photocoagulation (burn area approximately 30% of the treated retinal surface area). The theoretical basis for this approach is discussed. RESULTS: Untreated eyes had a preretinal PO2 of 22 +/- 9 mm Hg (mean +/- SD, n = 15 eyes). The preretinal PO2 was significantly higher over confluent, 12-day-old ALP or DLP lesions (51 +/- 13 mm Hg, n = 8 eyes; P < 0.01) compared to untreated eyes. However, at that time, DLP lesions had significantly higher PO2 values (60 +/- 13 mm Hg, n = 4 eyes) than did ALP lesions (42 +/- 6 mm Hg, n = 4 eyes; P = 0.04). The preretinal space in between laser lesions generally showed no significant increase in PO2 (P > 0.05) over controls on post-treatment days 1, 5, 14 and 47. The only exception was in the DLP group of eyes, in which a significant increase in PO2 over untreated or ALP-treated eyes occurred on post-treatment day 5 (41 +/- 7 mm Hg, n = 5 eyes; P = 0.01). Over photocoagulation lesions in this study, DLP produced a greater increase in preretinal PO2 compared to control values than did ALP. CONCLUSIONS: These results support the use of DLP as an alternative to ALP for the treatment of retinal vascular diseases in which hypoxia is suspected to play a role.

Role of dissolved plasma oxygen in hyperoxia-induced contrast.

Currently, hyperoxia is being investigated as a method for producing contrast in magnetic resonance images of the brain, solid tumors, and the eye. However, the underlying physiological mechanisms involved in this type of contrast are still not completely understood. For example, under what conditions would dissolved plasma oxygen contribute to the hyperoxia-induced contrast? Using the eye as a model system, we varied the level of dissolved plasma oxygen and observed different patterns of contrast in the vitreous. The observed contrast changes were consistent with tissue oxygen buffering by hemoglobin at an arterial PO2 of 200 mm Hg and dissolved oxygen offloading at arterial PO2's > 350 mm Hg. These data demonstrate that dissolved plasma oxygen does not become an important contrast mechanism until the arterial oxygen tension exceeds approximately 350 mm Hg. The implication of this result to studies in other organs is discussed.

Adult and newborn rat inner retinal oxygenation during carbogen and 100% oxygen breathing. Comparison using magnetic resonance imaging delta Po2 mapping.

PURPOSE: To test the hypothesis that breathing carbogen (95% O2-5% CO2) oxygenates the inner retina better than breathing 100% oxygen using an magnetic resonance imaging (MRI) method that noninvasively measures inner retinal oxygenation in normal adult and newborn rats. METHODS: Urethane-anesthetized adult and newborn (day 18) rats were studied. Sequential images were acquired in room air combined with either 100% oxygen or carbogen breathing. Normalized vitreous signal intensity changes were converted to oxygen tension changes (delta PO2) either on a pixel-by-pixel basis or in specific regions of interest. RESULTS: Systemic levels of hyperoxia during carbogen or 100% oxygen breathing were not significantly different (P > 0.05). In the adult rat, a significant difference (P = 0.017) was found in the preretinal vitreous delta PO2 during the breathing of either carbogen (130 +/- 9 mm Hg, mean +/- SEM; n = 5) or 100% oxygen (88 +/- 16 mm Hg; n = 5). Agreement was found between the MRI-determined delta PO2 values and literature oxygen microelectrodes data. In the newborn rat, significant differences (P < 0.05) in the preretinal vitreous delta PO2 were found during carbogen (164 +/- 23 mm Hg; n = 3) and oxygen breathing (91 +/- 8 mm Hg; n = 3). CONCLUSIONS: MRI delta PO2 mapping demonstrated for the first time that in the normal adult and newborn rat eye, carbogen breathing oxygenates the inner retina better than 100% oxygen breathing.

Hypoxia precedes the development of experimental preretinal neovascularization.

BACKGROUND: Although the mechanism of preretinal neovascular growth in the cell-injected rabbit eye model is not known, it has been proposed that the initial vasodilation and eventual development of neovascularization may be attributable to inflammatory mediators. However, an alternative explanation involving hypoxia has not been considered. The purpose of this study was to measure preretinal oxygen tension prior to the development of preretinal neovascularization in the cell-injected rabbit eye. METHODS: In the rabbit, intravitreous injections of 250,000 homologous dermal fibroblasts were performed on one eye; the fellow (control) eye was injected with vehicle. Preretinal oxygen tension over the myelin wing was measured using 19F-NMR spectroscopy of a 30-microliters droplet of perfluorocarbon previously injected into the preretinal vitreous. RESULTS: Compared to control eyes, fibroblast-injected eyes showed a 1.7-fold decrease in preretinal oxygen tension from the first time studied (1 day after cell injection) through the development of visible neovascularization. Hypoxia occurred without coexisting ophthalmoscopic evidence of vascular occlusion or, on days 1 and 3 after cell injection, retinal detachment. CONCLUSION: This result demonstrates for the first time that preretinal hypoxia precedes the development of preretinal neovascularization in the fibroblast-injected rabbit eye.