Retinal Blood Flow In Response Research Articles

Beneficial Effect of Long-Term Administration of Supplement With Trapa Bispinosa Roxb. and Lutein on Retinal Neurovascular Coupling in Type 2 Diabetic Mice.

PurposeWe investigated the effect of long-term administration of supplement with trapa bispinosa roxb. extract (TBE) and lutein on the susceptibility of retinal blood flow regulation in type 2 diabetic mice.MethodsSix-week-old db/db mice were randomly divided into the untreated group (n = 6) and the treated group received the supplement with TBE and lutein (n = 6). The longitudinal changes in retinal blood flow responses to systemic hyperoxia and a flicker stimulation were evaluated every 2 weeks in diabetes db/db mice from age 8 to 14 weeks. The retinal blood flow was assessed using laser speckle flowgraphy. We also evaluated the expressions of glial fibrillary acid protein (GFAP) and vascular endothelial growth factor (VEGF) by immunofluorescence.ResultsThe resting retinal blood flow was steady and comparable between two groups throughout the study. In db/db mice with supplement, both blood flow responses were restored from 8 to 14 weeks of age compared with diabetic mice treated with the placebo. Supplement prevented the activation of GFAP and decreased the expression of VEGF detected by immunofluorescence compared with the diabetic mice treated with placebo.ConclusionWe found that the long-term administration of supplement with TBE and lutein improved the impaired regulation of retinal blood flow in response to systemic hyperoxia and flicker stimulation, suggesting that these supplements can prevent diabetic retinopathy by improving abnormal neurovascular coupling in type 2 diabetic mice.

Longitudinal stability of retinal blood flow regulation in response to flicker stimulation and systemic hyperoxia in mice assessed with laser speckle flowgraphy

This study aimed to evaluate longitudinal changes in retinal blood flow in response to flicker stimulation and systemic hyperoxia in mice using a laser speckle flowgraphy (LSFG-Micro). The retinal blood flow in vascular area surrounding the optic nerve head was measured in 8-week-old male mice every 2 weeks until age 20-week. The coefficient of variation of retinal blood flow under resting condition was analyzed every 2 weeks to validate the consistency of the measurement. On day 1 of the experiment, retinal blood flow was assessed every 20 s for 6 min during and after 3 min flicker light (12 Hz) stimulation; on day 2, retinal blood flow was measured every minute for 20 min during and after 10 min systemic hyperoxia; and on day 3, electroretinography (ERG) was performed. Body weight, systemic blood pressure, and ocular perfusion pressure increased significantly with age, but the resting retinal blood flow and ERG parameters remained unchanged. Retinal blood flow significantly increased with flicker stimulation and decreased with systemic hyperoxia, independent of age. The LSFG-Micro provides consistent and reproducible retinal blood flow measurement in adult mice. Longitudinal assessments of retinal blood flow in response to flicker stimulation and systemic hyperoxia may be useful indexes for noninvasive monitoring of vascular function in retinas.

Changes in Retinal Blood Flow in Response to an Experimental Increase in IOP in Healthy Participants as Assessed With Doppler Optical Coherence Tomography.

PurposeBlood flow autoregulation is an intrinsic mechanism of the healthy retinal vasculature to keep blood flow constant when ocular perfusion pressure (OPP) is changed. In the present study, we set out to investigate retinal blood flow in response to an experimental decrease in OPP in healthy participants using Doppler optical coherence tomography.MethodsFifteen healthy participants aged between 22 and 31 years (mean, 27 ± 3 years) were included in the present open study. IOP was increased stepwise via the suction cup method to induce a decrease in OPP. Retinal blood flow in arteries and veins was assessed using a custom-built Doppler optical coherence tomography system and pressure–flow relationships were calculated to assess autoregulation.ResultsSuction cup application induced a pronounced increase in IOP with a maximum value of 50.5 ± 8.0 mm Hg at the highest level of suction. Pressure–flow relationships revealed that blood flow was autoregulated until the OPP was decreased by approximately 21 mm Hg and started to decrease significantly when the OPP was reduced by 30 mm Hg. Retinal blood flow at the last suction period decreased at a maximum of –57.0 ± 22.3% and 65.2 ± 15.4% in retinal arteries and retinal veins, respectively. These changes in retinal blood flow were less pronounced than the decrease in OPP (–75.2 ± 19.2%), indicating retinal autoregulation.ConclusionsThe results of the present study confirm that retinal blood flow is autoregulated in response to changes in the OPP. Doppler optical coherence tomography has the potential to become a clinical tool for the investigation of retinal blood flow autoregulation in the future, because of its ability to assess the blood velocities and diameter of the retinal vessels parallel and therefore also their blood flow in absolute values. (Clinicaltrials.gov number NCT03398616)

Role of glial cells in regulating retinal blood flow during flicker‐induced hyperemia and systemic hyperoxia‐induced hyporemia in cats

SummaryBackgroundTo investigate the role of glial cells in the regulation of retinal circulation in response to flicker stimulation, which is related to neurovascular coupling, and systemic hyperoxia in the retina.MethodsUsing laser Doppler velocimetry, we measured the retinal blood flow (RBF) in first‐order retinal arterioles in anesthetized cats. After intravitreal injection of l‐2‐aminoadipic acid (LAA) as gliotoxic compound, we examined the changes in RBF in response to flicker stimulation of 16 Hz for 3 min and systemic hyperoxia induced by the inhalation of 100% oxygen for 10 min.ResultsThe RBF increased gradually and reached a maximal level after 2–3 min of flicker stimuli in the PBS group. In the LAA‐treated eyes, the increases in RBF during the flicker stimulation were significantly attenuated by one third of those in the control eyes. During hyperoxia, the decreases in RBF in response to hyperoxia were significantly attenuated in the LAA‐treated eyes compared with the control.ConclusionsThe current results suggest that the retinal glial cells play an important role in the regulation of retinal blood flow during Flicker‐Induced Hyperemia and Systemic Hyperoxia‐Induced Hyporemia.

Role of Glial Cells in Regulating Retinal Blood Flow During Flicker-Induced Hyperemia in Cats

To investigate how glial cells participate in retinal circulation during flicker stimulation in cats. Using laser Doppler velocimetry, we measured the vessel diameter and blood velocity simultaneously and calculated the retinal blood flow (RBF) in feline first-order retinal arterioles. Twenty-four hours after intravitreal injections of L-2-aminoadipic acid (LAA), a gliotoxic compound, and the solvent of 0.01 N hydrochloric acid as a control, we examined the changes in RBF in response to 16-Hz flicker stimulation for 3 minutes. We also measured the changes in RBF 2 hours after intravitreal injection of Nω-propyl-L-arginine (L-NPA), a selective neuronal nitric oxide synthase inhibitor, in LAA-treated eyes. To evaluate the effects of LAA on retinal neuronal function, ERGs were monitored. Immunohistochemical examinations were performed. In LAA-treated eyes, histologic changes selectively occurred in retinal glial cells. There were no significant reductions in amplitude or elongation of implicit time in ERG after LAA injections compared with controls. In control eyes, the RBF gradually increased and reached the maximal level (53.5% ± 2.5% increase from baseline) after 2 to 3 minutes of flicker stimulation. In LAA-treated eyes, the increases in RBF during flicker stimulation were attenuated significantly compared with controls. In LAA-treated eyes 2 hours after injection of L-NPA, flicker-evoked increases in RBF decreased significantly compared with LAA-treated eyes. The current results suggested that increases in RBF in response to flicker stimulation were regulated partly by retinal glial cells.

Optical Coherence Tomography Angiography of Peripapillary Retinal Blood Flow Response to Hyperoxia.

To measure the change in peripapillary retinal blood flow in response to hyperoxia by using optical coherence tomography (OCT) angiography. One eye of each healthy human participants (six) was scanned with a commercial high-speed (70 kHz) spectral OCT. Scans were captured twice after 10-minute exposures to normal breathing (baseline) and hyperoxia. Blood flow was detected by the split-spectrum amplitude-decorrelation angiography (SSADA) algorithm. Peripapillary retinal blood flow index and vessel density were calculated from en face maximum projections of the retinal layers. The experiment was performed on 2 separate days for each participant. Coefficient of variation (CV) was used to measure within-day repeatability and between-day reproducibility. Paired t-tests were used to compare means of baseline and hyperoxic peripapillary retinal blood flow. A decrease of 8.87% ± 3.09% (mean ± standard deviation) in flow index and 2.61% ± 1.50% in vessel density was observed under hyperoxia. The within-day repeatability CV of baseline measurements was 5.75% for flow index and 1.67% for vessel density. The between-day reproducibility CV for baseline flow index and vessel density was 11.1% and 1.14%, respectively. The between-day reproducibility of the hyperoxic response was 3.71% and 1.67% for flow index and vessel density, respectively. Optical coherence tomography angiography with SSADA was able to detect a decrease in peripapillary retinal blood flow in response to hyperoxia. The response was larger than the variability of baseline measurements. The magnitude of an individual's hyperoxic response was highly variable between days. Thus, reliable assessment may require averaging multiple measurements.

Role of neuronal nitric oxide synthase in regulating retinal blood flow during flicker-induced hyperemia in cats.

To investigate how neuronal nitric oxide synthase (nNOS) contributes to regulation of the retinal circulation during rest and flicker stimulation in cats. Using laser Doppler velocimetry, we measured the vessel diameter and blood velocity simultaneously and calculated the retinal blood flow (RBF) in feline first-order retinal arterioles. After intravitreal injections of Nω-Nitro-L-arginine methyl ester (L-NAME), a nonselective NOS inhibitor, and Nω-propyl-L-arginine (L-NPA), a selective nNOS inhibitor, we continuously monitored the retinal circulation without any perturbations for 2 hours. We then examined the changes in the RBF in response to 16-Hz flicker stimuli for 3 minutes at 2 hours after intravitreal injection of phosphate-buffered saline (PBS) as a control, L-NAME, L-NPA, and thromboxane A2 (TXA2) analogue U46619 as a basal tone-adjusted control. After intravitreal injection of L-NAME and L-NPA, the baseline RBF decreased gradually in a dose-dependent manner. In the PBS group, the RBF increased gradually and reached a maximal level after 2 to 3 minutes of flicker stimuli. After 3 minutes of 16-Hz flicker stimuli, the RBF increased by 53.5% ± 3.4% compared with baseline. In the L-NAME and L-NPA groups, the increases in RBF during flicker stimulation were attenuated significantly compared with the PBS group. In the TXA2 group, the reduction in the flicker-induced increase in RBF was comparable to that in the PBS group. The current results suggested that increased RBF in response to flicker stimulation may be mediated by nitric oxide (NO) production via nNOS activation.

Autoregulation of Retinal Blood Flow in Response to Decreased Ocular Perfusion Pressure in Cats: Comparison of the Effects of Increased Intraocular Pressure and Systemic Hypotension

To investigate the regulatory mechanisms responsible for autoregulation of retinal blood flow (RBF) during periods of decreased ocular perfusion pressure (OPP). The effects of acute reductions in OPP on RBF were assessed using laser Doppler velocimetry in cats. The OPP decreased from 90 to 40 mm Hg by increasing the IOP (elevated IOP) or by decreasing the systemic blood pressure via exsanguination (systemic hypotension). The contributions of nitric oxide (NO), adenosine, and/or N-methyl-D-aspartic acid (NMDA) in regulation of the retinal arteriolar hemodynamics during decreased OPP was determined at 120 minutes after intravitreal injection of various inhibitors or PBS. Following PBS injection, the flow velocity decreased in proportion to the decrease in OPP; however, the retinal arteriolar diameter gradually increased. Consequently, the RBF was maintained near baseline levels when the OPP exceeded 70 mm Hg but decreased significantly (P < 0.01) when the OPP fell to less than or equal to 60 mm Hg due to elevated IOP or systemic hypotension. Adenosine receptor blocker 8-(p-sulfophenyl)theophylline, significantly (P < 0.01) enhanced decreases in RBF induced by elevated IOP and systemic hypotension at OPP from 80 to 40 mm Hg, whereas NO synthase inhibitor N(G)-nitro-L-arginine-methyl ester and NMDA receptor antagonist DL-2-amino-5-phosphonopentanoic acid only significantly (P < 0.01) enhanced reductions in RBF induced by elevated IOP. These results indicate that adenosine contributes to autoregulation of RBF during systemic hypotension, whereas adenosine, NO, and NMDA receptors autoregulate the RBF after elevated IOP. Different vasoregulatory factors might contribute to autoregulation of RBF after decreases in OPP induced by elevated IOP and systemic hypotension.

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Effects of Dorzolamide-Timolol and Brimonidine-Timolol on Retinal Vascular Autoregulation and Ocular Perfusion Pressure in Primary Open Angle Glaucoma

To assess whether dorzolamide 2%-timolol 0.5% (D/T) and/or brimonidine 0.2%-timolol 0.5% (B/T) alters retinal vascular autoregulation (RVA) and seated ocular perfusion pressure (sOPP) in primary open angle glaucoma (POAG) patients who demonstrate retinal vascular dysregulation (RVD) on timolol 0.5% alone. In this prospective, observer-masked, crossover study, 21 POAG patients with untreated intraocular pressure (IOP) >21 mmHg were treated for 6 weeks with timolol 0.5%. Subsequently, we measured inferior temporal retinal artery blood flow in the left eye with subjects seated and then while reclined for 30 min using the Canon Laser Blood Flowmeter. Subjects with a change in retinal blood flow in response to posture change outside of the range previously found in healthy subjects were designated as having RVD and randomized to either D/T or B/T for 6 weeks and re-tested. This was followed by treatment with the opposite medication. Seven of the 21 subjects demonstrated RVD in response to posture change following timolol 0.5%. Multiple linear regression analysis indicated that lower sOPP was the main determinant of RVD (P=0.033). After treatment with D/T, all 7 converted from RVD to normal RVA status (P=0.001). Four of 6 subjects showed a similar return to normal RVA following B/T (P=0.066). Mid-morning sOPP was 41.1±5.5 mmHg post-timolol, 46.3±6.5 mmHg post-D/T, and 38.6±6.0 mmHg post-B/T (D/T vs. B/T, P=0.026). D/T significantly improved RVA in POAG patients exhibiting RVD while on timolol 0.5% alone. D/T also increased sOPP compared to B/T. There was no significant difference (P=0.37) between D/T and B/T in improving RVA.

Retinal blood flow in response to an intravitreal injection of ranibizumab for neovascular age‐related macular degeneration

To assess the hemodynamic response of retinal arterioles and venules following a single intravitreal injection of ranibizumab in neovascular age-related macular degeneration (NV-AMD) patients and to assess the influence of the number of prior injections on this response. Fifteen NV-AMD patients were prospectively recruited and grouped according to the dosage of ranibizumab previously received. Group 1 NV-AMD patients (n = 7) had previously received 1.50 mg or less, and group 2 patients (n = 8) had received more than 1.50 mg in the study eye. A group of 12 non-NV AMD patients were also recruited for control comparison. Vessel diameter, centreline blood velocity and blood flow were assessed with the Canon Laser Blood Flowmeter immediately prior to an injection and at a mean follow-up of 37.7 and 36.7 days for group 1 and group 2 patients, respectively. The NV-AMD patients as a whole and the group 1 cohort had a significantly greater arteriolar diameter at baseline than the non-NV AMD patients. There was a significant reduction in arteriolar diameter, velocity and blood flow in group 1 but not in group 2 NV-AMD patients at follow-up. There was only an insignificant decrease in measured parameters of the retinal venules. At follow-up, there was no difference in the diameter, velocity or flow between AMD patients. Intravitreal ranibizumab treatment for NV-AMD induces a reduction in arteriolar diameter, velocity, and blood flow in patients who have received <1.50 mg of ranibizumab.

Role of Nitric Oxide in Regulation of Retinal Blood Flow in Response to Hyperoxia in Cats

To investigate whether nitric oxide (NO) regulates retinal circulation during and after induction of hyperoxia in cats. Hyperoxia was induced for 10 minutes with 100% oxygen. The vessel diameter and blood velocity were measured simultaneously in second-order retinal arterioles by laser Doppler velocimetry; the retinal blood flow (RBF) and wall shear rate (WSR) were calculated during and after hyperoxia. PBS, L-NAME, D-NAME, BQ-123, BQ-788, and 7-nitroindazole (7-NI) were administered before induction of hyperoxia. In the PBS group, vessel diameter, blood velocity, and RBF decreased during hyperoxia and returned to baseline within 10 minutes after hyperoxia ended. WSR decreased transiently and then returned to baseline by the delayed constriction of retinal arterioles during hyperoxia. In the l-NAME and BQ-788 groups, the decreases in RBF during hyperoxia did not differ from those in the PBS group. However, the recovery of RBF after hyperoxia ended was attenuated significantly until 20 minutes after hyperoxia ended in both groups compared with the PBS group (P < 0.05). In the BQ-123 group, the intravitreous injection of BQ-123 caused less reduction of blood velocity and RBF during hyperoxia compared with that in the PBS group, whereas the RBF immediately returned to baseline after hyperoxia. D-NAME and 7-NI did not affect RBF in response to hyperoxia. The current results indicate that NO contributes to RBF recovery after hyperoxia, probably through the action of endothelial NOS via the ETB receptor in the vascular endothelium of the retinal arterioles, suggesting that the RBF response to hyperoxia may be used to evaluate the endothelial function of the retinal arterioles.

Mechanisms of Hyperoxia-Induced Reductions in Retinal Blood Flow in Newborn Pig

Although reductions in retinal blood flow (RBF) in response to acute hyperoxia are well described, the mechanistic basis of this response has yet to be clarified. The present study was undertaken in order to determine the possible involvement of two arachidonic acid-derived vasoconstrictors, the cyclooxygenase metabolite thromboxane and the cytochrome P450metabolite 20-HETE, as well as the involvement of the peptide endothelin and superoxide free radical. Fluorescein videoangiography was performed on the intact eyes of isoflurane-anesthetized newborn piglets. RBF responses to 20 min of hyperoxia were calculated from the angiograms off-line, using changes in mean arteriovenous transit times and arteriolar and venular diameters. The effect of hyperoxia (PaO2=351±9 mmHg;n=39) on RBF was examined in each animal under control conditions and again after intravitreal perivascular administration of drugs that block the synthesis or receptors of known vasoconstrictors. Estimated RBF decreased by a maximum of 42±3% in the 7 animal groups in response to 20 min of hyperoxia. The magnitude and time course of the change in RBF resulting from two successive hyperoxic challenges did not differ, and were unaffected by intravitreal administration of vehicle. The response to hyperoxia was attenuated 46±6 (n=6;P=0.001) after intravitreal CGS 22652 (2 nmol), a combined thromboxane synthesis inhibitor and receptor antagonist. DDMS (12.5 nmol), a competitive inhibitor of the P450enzyme ω-hydroxylase that forms 20-HETE, blocked hyperoxic constriction by 23±7% (n=6;P=0.01). Intravitreal pretreatment with TBC 1241z (2 nmol), a receptor antagonist of the peptide endothelin, blocked the hyperoxic response by 26±5% (n=6;P=0.01). A combination of CGS 22652 (2 nmol), DDMS (12.5 nmol), and TBC 1241z (2 nmol), blocked the hyperoxic flow response by 51±3% (n=5;P=0.003). Administration of a combination of superoxide dismutase (10 U intravitreally, 10000 U kg-1of the polyethylene glycol-conjugate intravenously) and catalase (10 U intravitreally, 10000 U kg-1intravenously) was without effect on hyperoxia-induced reductions in RBF (n=5). The present results indicate that the arachidonic acid metabolites thromboxane and 20-HETE, and the peptide endothelin, participate in mediating the acute reduction in RBF in response to hyperoxia.

Physiological perturbation of ocular and cerebral blood flow as measured by scanning laser ophthalmoscopy and color doppler imaging

Retinal blood flow regulation in health remains poorly described. We hypothesized that retinal perfusion is controlled to provide constant O 2 delivery to that tissue, and that changes in retinal blood flow in response to chemical stimuli parallel changes in carotid and retrobulbar perfusion. Accordingly, in 11 young adults with normal eye examinations, we measured retinal blood flow indices (via scanning laser ophthalmoscopy [SLO] during fluorescein angiography) and carotid, ophthalmic, and central retinal arterial blood flow indices (via Doppler imaging [CDI]) under control, hypoxic (alveolar PO 2 = 55 ± 3 mmHg) and hyperoxic (alveolar PO 2 = 655 ± 18 mmHg) conditions. The three conditions were counterbalanced in order and isocapnia was maintained in each. Retinal arterial mean dye velocity and arteriovenous passage time, as measured by SLO, were slowed by hyperoxia and accelerated by hypoxia, in rough proportion to the changes in arterial O 2 content (± 10%; p < 0.05). In the seven subjects in which relative measurements of retinal arterial diameters were obtained, neither hypoxia nor hyperoxia significantly altered vessel diameter. At the same time, mean retinal capillary transit velocity was independent of PO 2, suggesting that, in health, retinal capillaries may be recruited as PO 2 falls. O 2-induced changes in carotid, ophthalmic, or central retinal arterial blood flow velocities (via CDI) were not found, though a wide coefficient of variation (30% for CDI vs. 14% for SLO) may have contributed to this failure. We conclude that, under isocapnic conditions, retinal perfusion may be regulated to provide constant O 2 delivery. The lack of precision of CDI in this study leaves open the possiblity that chemical regulation of the retinal flow may parallel that of other brain regions.

Retinal laser Doppler velocimetry: toward its computer-assisted clinical use

Clinical use of laser Doppler velocimetry of absolute red blood cell velocity V(max) in the center of major retinal vessels becomes possible with recent advances in microcomputer technology- Speed and automation of Doppler photocurrent analysis and identification of proper laser beam positioning are the major requirements in making direct quantitative assessment of retinal hemodynamics a routine diagnostic tool. We discuss our efforts toward achieving this goal and illustrate our current capabilities with examples of changes in retinal blood flow in response to physiologic maneuvers. In veins, V(max) can now be determined on-line. In arteries, current computing speed only supports intermittent on-line data acquisition.