OBJECTIVEIncreased vascular endothelial growth factor (VEGF) and VEGF receptor expression is the important biological response under shear stress, ischemia and hypoxia conditions. Mechanical vibration induced cochlea shear stress and trauma obviously upregulate VEGF and VEGF receptor 2 (VEGFR2) expression in the cochlea. To evaluate the possibility of VEGF varying the transport in blood-labyrinth barrier and blood-perilymphatic barrier.

METHODSEleven guinea pigs, male and female, weighing from 300 g to 900 g were kept under general anaesthesia with xylazine (16 mg/kg) and ketamine (60 mg/kg) for both drug delivery and MRI measurement. VEGF (6 ears) and phosphate-buffered saline (PBS, 5 ears) were delivered to the inner ear via the round window membrane (soaked in gelfoam). The T1 contrast agent gadodiamide (Gd-DTPA-BMA) chelated bound paramagnetic gadolinium was used as the inner ear barrier transportation tracer. A Bruker Biospec Avance 47/40 experimental MRI system with a magnetic field strength of 4. 7 Tesla and a 40 cm bore was used for the 2-dimensional cochlea MRI evaluation. The Paravision software was used for image intensity measurement and the Adobe Photoshop 6.0 software was used for image presentation.

RESULTSVEGF induced significant Gd uptake in the scala tympani and scala vestibuli, but had little effect on the uptake of Gd in the scala media.

CONCLUSIONSVEGF significantly increased the transportation of blood-perilymphatic barrier and adapted the inner ear for compensation and repair.

Animals ; Blood-Brain Barrier ; drug effects ; Blood-Retinal Barrier ; drug effects ; Ear, Inner ; drug effects ; metabolism ; Female ; Guinea Pigs ; Magnetic Resonance Imaging ; Male ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVE: The purpose of this study is to evaluate the effect of dexamethasone on the damaged blood-ocular barrier caused by triolein emulsion, using contrast-enhanced MR imaging. MATERIALS AND METHODS: An emulsion of 0.1-mL triolein in 20 mL of saline was infused into the carotid arteries of 32 cats, 12 cats were placed in the treatment group and 18 cats were placed in the Control group. Thirty minutes after the infusion of triolein emulsion, a set of orbital pre- and post-contrast T1-weighted MR images (T1WIs) were obtained. Infusion of 10 mg/kg dexamethasone into the ipsilateral carotid artery of each of the cats in the treatment group cats and 20 mL saline in each of the cats in the control group was given. A second set of pre- and post-contrast orbital T1WIs were obtained three hours following triolein emulsion infusion. Qualitative analysis was performed for the the anterior chamber (AC), the posterior chamber (PC), and in the vitreous humor of the ipsilateral and contralateral eyes. The signal intensity ratios of the ipsilateral eye over the contralateral eye were quantitatively evaluated in the three ocular chambers on the first and second set of T1WIs, and were then statistically compared. RESULTS: Qualitatively, the AC, the PC or the vitreous did not show immediate contrast enhancement on the first and the second set of post-contrast T1WIs. However, the AC and the PC showed delayed contrast enhancement for both groups of cats on the second pre-contrast T1WIs. No enhancement or minimally delayed enhancement was seen for the vitreous humor. Quantitatively, the signal intensity ratios in the PC of the treatment group of cats were statistically lower than the ratios of the control group of cats for the second set of T1WIs (p = 0.037). The AC and vitreous showed no statistically significant difference between the feline treatment group and control group (p > 0.05). CONCLUSION: Contrast-enhanced MR images revealed increased vascular permeability in the PC of the eye after infusion of triolein emulsion. Dexamethasone seems to decrease the breakdown of the blood-aqueous barrier in the PC.
Animals ; Blood-Aqueous Barrier/*drug effects ; Blood-Retinal Barrier/*drug effects ; Capillary Permeability/drug effects ; Cats ; Contrast Media ; Dexamethasone/*pharmacology ; Emulsions ; Glucocorticoids/*pharmacology ; Image Enhancement ; Magnetic Resonance Imaging/*methods ; Triolein/*adverse effects

PURPOSE: Cancer-associated retinopathy is a paraneoplastic retinal degeneration which may primarily result from auto-immune mediated apoptosis. It has been hypothesized that high titer of auto-antibodies are able to cross the blood-retinal barrier (BRB) and to enter retinal cells to activate apoptotic pathway which has been already well-established. However, it still remains to be elucidated whether auto-antibodies could cross BRB in the retina. Herein, we demonstrated that intravenously administrated anti-recoverin antibodies could not pass through BRB and not lead to retinal cell death. METHODS: Anti-recoverin antibody was intravenously injected to C57BL/6 mice, which were sacrificed 1 and 7 days to obtain eye. Vascular endothelial growth factor was intravitreally injected to induce BRB breakdown, which was confirmed by fluorescein angiography and western blotting for zonula occludens (ZO)-1, ZO-2 and occludin. To investigate the location of anti-recoverin antibody in the retina, immunofluorescein was performed. The retinal toxicity of intravenous anti-recoverin antibody was evaluated by histological examination and transferase-mediated dUTP nick-end labeling. Immunofluorescein staining for glial fibrillary acidic protein was done to address glial activation as well. RESULTS: Intravenously administrated anti-recoverin antibodies were exclusively distributed on retinal vessels which were co-localized with CD31, and led to neither increase of glial fibrillary acidic protein expression, as an indicator of retinal stress, nor apoptotic retinal cell death. Moreover, even in the condition of vascular endothelial growth factor-induced BRB breakdown, anti-recoverin antibodies could not migrate across BRB and still remained on retinal vessels without retinal cytotoxicity. CONCLUSIONS: Our results suggest that high titer of intravascular anti-recoverin antibodies could not penetrate into the retina by themselves, and BRB breakdown mediated by dysregulation of tight junction might not be sufficient to allow anti-recoverin antibodies to pass through BRB.
Animals ; Antibodies/*administration & dosage/*metabolism ; Blood-Retinal Barrier/*metabolism ; Cell Death/drug effects ; Cells, Cultured ; Female ; Injections, Intravenous ; Mice ; Mice, Inbred C57BL ; Recoverin/*immunology ; Retina/cytology/drug effects ; Retinal Vessels/metabolism

Using computerized vitreous fluorophotometry (VFP, Fluorotron(TM)), we examined the effect of cryotherapy on the blood retinal barrier (BRB) and the effect of subtenon injection of methylprednisolone acetate (Depomedrol(R)). In experiment 1, the right eyes of the 13 pigmented rabbits were treated with heavy cryotherapy after baseline VFP readings. The freezes were applied at 6 places in each quadrant around the equator are in two rows, a total of 24 places circumferentially. The left eyes were reserved as controls. In 6 rabbits (cryo with steroid group), Depomedrol(R) 10 mg of Depomedrol was injected into subtenon space after cryotherapy. The other 7 rabbits were treated with cryotherapy only (cryo only group). The VFP readings were taken 1, 3, 5, and 7 days, 2, 3, 5, and 7 weeks after cryotherapy. Cryotherapy increased the breakdown of BRB significantly. The peak VFP readings were obtained 5 days after cryotherapy in the cryo only group and 7 days after cryotherapy in the cryo with steroid group. In the cryo only group, the severity of the breakdown of BRB was higher than in the cryo with steroid group, and the increased VFP readings could not be normalized until 7 weeks after cryotherapy. In experiment 2, both eyes of the 8 pigmented rabbits were treated with medium cryotherapy after baseline VFP readings. The freezes were applied at 3 places in the superior temporal quadrant and at 3 places in the superior nasal quadrant, a total of 6 places. Depomedrol(R) 10 mg was injected into subtenon space after cryotherapy in the right eyes only. The VFP readings were taken 1, 3, 5, 7, 10, and 14 days after cryotherapy. In this experiment, cryotherapy did not increase the breakdown of BRB. But in the right eye, the severity of the breakdown of BRB was significantly lower than in the left eye 7 and 10 days after cryotherapy. These results suggest that Depomedrol(R) can decrease the severity of the breakdown of BRB after cryotherapy, and may be useful in the prevention of proliferative vitreoretinopathy (PVR).
Animals ; Anti-Inflammatory Agents/*pharmacology ; Blood-Retinal Barrier/*drug effects ; Capillary Permeability ; *Cryosurgery/adverse effects ; Fascia ; Fluorophotometry ; Injections ; Methylprednisolone/*analogs & derivatives/pharmacology ; Orbit ; Rabbits ; Retina/*drug effects/surgery ; Vitreoretinopathy, Proliferative/prevention & control

To evaluate the effect of chlorogenic acid (CGA), a polyphenol abundant in coffee, on retinal vascular leakage in the rat model of diabetic retinopathy, Sprague-Dawley rats were divided into four groups: controls, streptozotocin-induced diabetic rats, and diabetic rats treated with 10 and 20 mg/kg chlorogenic acid intraperitoneally daily for 14 days, respectively. Blood-retinal barrier (BRB) breakdown was evaluated using FITC-dextran. Vascular endothelial growth factor (VEGF) distribution and expression level was evaluated with immunohistochemistry and Western blot analysis. Expression of tight junction proteins, occludin and claudin-5, and zonula occludens protein, ZO-1 was also evaluated with immunohistochemistry and Western blot analysis. BRB breakdown and increased vascular leakage was found in diabetic rats, with increased VEGF expression and down-regulation of occludin, claudin-5, and ZO-1. CGA treatment effectively preserved the expression of occludin, and decreased VEGF levels, leading to less BRB breakdown and less vascular leakage. CGA may have a preventive role in BRB breakdown in diabetic retinopathy by preserving tight junction protein levels and low VEGF levels.
Animals ; Blood-Retinal Barrier/*drug effects ; Chlorogenic Acid/metabolism/*pharmacology ; Claudin-5/metabolism ; Dextrans/chemistry ; Diabetes Mellitus, Experimental/complications/metabolism/*pathology ; Diabetic Retinopathy/etiology/prevention & control ; Down-Regulation ; Fluorescein-5-isothiocyanate/chemistry ; Male ; Occludin/metabolism ; Rats ; Rats, Sprague-Dawley ; Retina/*metabolism ; Tight Junction Proteins/metabolism ; Vascular Endothelial Growth Factor A/metabolism ; Zonula Occludens-1 Protein/metabolism