Blood flow rate estimation in optic disc capillaries and vessels using Doppler optical coherence tomography with 3D fast phase unwrapping

The retinal volumetric flow rate contains useful information not only for ophthalmology but also for the diagnosis of common civilization diseases such as diabetes, Alzheimer's disease, or cerebrovascular diseases. Non-invasive optical methods for quantitative flow assessment, such as Doppler optical coherence tomography (OCT), have certain limitations. One is the phase wrapping that makes simultaneous calculations of the flow in all human retinal vessels impossible due to a very large span of flow velocities. We demonstrate that three-dimensional Doppler OCT combined with three-dimensional four Fourier transform fast phase unwrapping (3D 4FT FPU) allows for the calculation of the volumetric blood flow rate in real-time by the implementation of the algorithms in a graphics processing unit (GPU). The additive character of the flow at the furcations is proven using a microfluidic device with controlled flow rates as well as in the retinal veins bifurcations imaged in the optic disc area of five healthy volunteers. We show values of blood flow rates calculated for retinal capillaries and vessels with diameters in the range of 12-150 µm. The potential of quantitative measurement of retinal blood flow volume includes noninvasive detection of carotid artery stenosis or occlusion, measuring vascular reactivity and evaluation of vessel wall stiffness

Influence of cardiovascular condition on retinal and retinal nerve fiber layer measurements

Objective To assess changes in the retinal nerve fiber layer (RNFL) and macula in subjects with cardiovascular risk factors or subclinical ischemia. Design Prospective and observational study. Methods A total of 152 healthy men underwent cardiovascular examination, including quantification of subclinical atheroma plaques by artery ultrasound scans, blood analysis, and a complete ophthalmic evaluation, including spectral-domain optical coherence tomography. The variables registered in cardiovascular examination were quantification of classic major risk factors, subclinical atheroma plaques by artery ultrasound scans, and analytical records. The ophthalmic evaluation registered RNFL and macular thickness. Results Mean subject age was 51.27±3.71 years. The 40 subjects without classic cardiovascular risk factors did not show differences in RNFL and macular thicknesses compared with the 112 subjects with at least one risk factor (except in sector 9 that showed higher thicknesses in subjects with 1 risk factor). Comparison between the group of subjects with and without atheroma plaques revealed no differences in RNFL and macular thicknesses. The sub-analysis of subjects with subclinical atheroma plaques in the common carotid artery revealed a significant reduction in central macular thickness in the left eye compared with the right eye (p = 0.016), RNFL in the superior quadrant (p = 0.007), and the 11 o’clock sector (p = 0.020). Comparison between smokers and nonsmokers revealed that smokers had significant thinning of the central macular thickness (p = 0.034), the nasal RNFL quadrant (p = 0.006), and the 3 and 5 o’clock sectors (p = 0.016 and 0.009). Conclusions Classic cardiovascular risk factors do not cause RNFL or macular thickness reduction, but tobacco smoking habit reduces nasal RNFL thickness. Subclinical atherosclerosis in the common carotid artery associates a reduction in central macular and nasal RNFL quadrant thicknesses in the left eye compared with the right eye

Measurement of cerebral microvascular compliance in a model of atherosclerosis with optical coherence tomography

Optical coherence tomography (OCT) has recently been used to produce 3D angiography of microvasculature and blood flow maps of large vessels in the rodent brain in-vivo. However, use of this optical method for the study of cerebrovascular disease has not been fully explored. Recent developments in neurodegenerative diseases has linked common cardiovascular risk factors to neurodegenerative risk factors hinting at a vascular hypothesis for the development of the latter. Tools for studying cerebral blood flow and the myogenic tone of cerebral vasculature have thus far been either highly invasive or required ex-vivo preparations therefore not preserving the delicate in-vivo conditions. We propose a novel technique for reconstructing the flow profile over a single cardiac cycle in order to evaluate flow pulsatility and vessel compliance. A vascular model is used to simulate changes in vascular compliance and interpret OCT results. Comparison between atherosclerotic and wild type mice show a trend towards increased compliance in the smaller arterioles of the brain (diameter < 80μm) in the disease model. These results are consistent with previously published ex-vivo work confirming the ability of OCT to investigate vascular dysfunction

Développement des systèmes d’imagerie basés sur la tomographie par cohérence optique visant l’étude des maladies cardiovasculaires

Les maladies cardiovasculaires (CVD) sont la principale cause de décès depuis des décennies. Son taux de morbidité élevé engendre un fardeau social et économique énorme et à la société. L’imagerie biomédicale est un outil important pour étudier et évaluer les CVDs. Notamment, la tomographie par cohérence optique (OCT) a montré des avantages attrayants pour la recherche appliquée aux CVDs. Le but global de cette thèse est le développement de systèmes OCT combinés avec d’autres techniques d’imagerie optique pour étudier les CVDs. Le premier objectif est de développer un système d'imagerie intravasculaire combinant l’OCT et l'imagerie par fluorescence proche infrarouge (NIRF), qui pourrait à terme être utilisé pour la détection et l'évaluation de l'athérosclérose dans les artères coronaires. L’originalité du travail se situe dans l’utilisation d’une nouvelle technique de détection de photons, la détection de photons à déclenchement rapide, intégrée dans notre système d’imagerie, ce qui a considérablement amélioré le rapport signal sur bruit et la sensibilité en profondeur de la NIRF. Une expérience ex vivo dans des conditions réalistes a validé le mécanisme de notre système d'imagerie intravasculaire à double modalité au niveau des aspects optique, mécanique et logiciel. Les images hybrides provenant de l'OCT et de la NIRF ont fourni des informations structurelles et moléculaires sur le fantôme imitant le vaisseau sanguin, ce qui suggère un grand potentiel d’utilisation de notre système d'imagerie chez des modèles animaux. Le deuxième objectif de cette thèse était d’étudier les impacts de l’athérosclérose sur différents aspects du cerveau de souris avec l’aide de plusieurs techniques d’imagerie optique, y compris l’imagerie intrinsèque optique, l’OCT et la microscopie à deux-photon. En comparaison avec les souris athérosclérotiques jeunes, le groupe âgé a montré un changement de concentration plus faible en hémoglobine oxygénée, hémoglobine désoxygénée et hémoglobine totale dans le cortex somatosensoriel à la suite de la stimulation par vibrisse, ce qui indique que la maladie d’athérosclérose réduit la réponse hémodynamique à la stimulation sensorielle. Les résultats obtenus à partir des données Doppler OCT ont révélé que le diamètre et le débit sanguin moyen des artérioles descendantes chez les souris ATX âgées étaient significativement plus petits comparés avec ceux des souris ATX jeunes, ce qui suggère que l'athérosclérose entraîne une dégénérescence structurelle et fonctionnelle des artérioles. L’altération fonctionnelle a également été observée dans les capillaires chez les souris ATX âgées, caractérisée par une plus faible vitesse es globules rouges (RBC), un plus faible flux de RBC, un plus faible hématocrite et une plus grande hétérogénéité du temps de transit capillaire. L’oxygénation tissulaire évaluée par la microscopie à deux photons a confirmé que l’altération de la microvascularisation liée à l’athérosclérose compromettait l’apport d’oxygène au tissu cérébral, amenant une hypoxie cérébrale chez les vieilles souris ATX. Les capillaires chez les souris ATX âgées ont été trouvés dilatés, ce qui pourrait être un mécanisme de régulation servant à compenser partiellement la diminution du débit sanguin associée à l'athérosclérose. Le troisième objectif de cette thèse est d’étudier la réparation du tissu cérébral et la régénération microvasculaire suite à un accident vasculaire cérébral (AVC) ischémique chez des souris en utilisant un système OCT construit dans le laboratoire. Le modèle d'AVC ischémique a été créé par occlusion photo-thrombotique des capillaires. Sur les images de l'OCT, seuls les tissus cérébraux profonds ont été endommagés par l'AVC ischémique, alors que les tissus proches de la surface corticale étaient intacts bien qu'ils aient été exposés à une intensité de laser plus élevée pendant la photo-thrombose. Cette observation implique que les tissus cérébraux profonds sont plus vulnérables lorsque l’apport d’oxygène des capillaires est interrompu au cours d’un AVC ischémique. Au cours de la récupération post-AVC, la lésion ischémique diminuait pendant que la régénération microvasculaire progressait. De plus, le réseau capillaire nouvellement formé autour de la lésion avait une structure hautement organisée et directionnelle avec un grand pourcentage de segments capillaires s'étendant vers le centre ischémique. Plus le réseau capillaire était profond, mieux sa structure était organisée, ce qui suggère que l'oxygénation des tissus profonds est plus dépendante du réseau capillaire. Notre analyse des événements de blocage capillaire a révélé que l’AVC ischémique augmentait à la fois la densité et l’incidence de blocage capillaire. Étant donné que le blocage capillaire pourrait entraîner une réduction du débit sanguin cérébral, il pourrait être utile de le cibler pour chercher de nouveaux traitements pour l'AVC.----------ABSTRACT Cardiovascular diseases (CVD) have been the leading cause of death for decades, and their high morbidity rate has led to enormous social and economic burden to both patients and society. Biomedical imaging systems are important tools to study and assess CVDs. Optical coherence tomography (OCT) has shown appealing advantages in CVD research. The general purpose of this thesis is the development of OCT-based imaging systems coupled with other optical imaging techniques to investigate CVDs. The first objective of this thesis is to develop an intravascular imaging system combining OCT and near infrared fluorescence (NIRF) imaging, which could ultimately be used for the detection and evaluation of atherosclerosis in coronary arteries. A novel photon detection technique, fast time-gating photon detection, was integrated into our imaging system, which improved significantly the signal-to-noise ratio and the depth sensitivity of NIRF. Ex vivo experiment under realist conditions validated the mechanism of our dual modality intravascular imaging system in terms of the optical, mechanical and software aspects. The hybrid images from OCT and NIRF provided structural and molecular information on the vessel-mimicking phantom, suggesting a great potential of our imaging system to be used in animal models. The second objective of this thesis is to study the impact of atherosclerotic disease on different aspects of mouse brain by using a variety of high-resolution optical imaging techniques, including intrinsic optical signal imaging, OCT and two-photon microscopy. Compared with the young atherosclerotic (ATX) mice, the old ones displayed a smaller change in the concentration of oxygenated hemoglobin, deoxygenated hemoglobin and total hemoglobin in the barrel cortex following whisker stimulation, which indicates that atherosclerotic disease is associated with reduced hemodynamic response to sensory stimulation. Results derived from Doppler OCT data revealed that the diameter and the mean blood flow of diving arterioles in the old ATX mice were significantly smaller compared with those in the young ATX mice, suggesting that atherosclerosis leads to structural and functional degeneration in descending arterioles. Functional alteration was also observed in capillaries among the old ATX mice with lower red blood cell (RBC) speed, lower RBC flux, lower hematocrit and higher transit time heterogeneity. Tissue oxygenation assessed by two-photon microscopy confirmed that atherosclerosis-related microvasculature impairment severely compromised oxygen supply to brain tissue, causing cerebral hypoxia in the old ATX mice. Capillaries in the old ATX mice were found dilated, which could be a regulatory mechanism of partially compensating atherosclerosis-associated blood flow decrease. The third objective of this thesis is to study post-stroke tissue repair and microvasculature regeneration and function in a mouse model of stroke using a custom-made OCT imaging system. Ischemic stroke model was created by photo-thrombotic occlusion of capillaries. On OCT structural and angiographic images, only deep cerebral tissue was found damaged by the ischemic stroke, whereas tissue close to the cortical surface seemed intact although it was exposed to a high laser intensity during the photo-thrombosis. This observation implies that deep cerebral tissue is more vulnerable when oxygen supply from capillaries are interrupted during ischemic stroke. Over the course of post-stroke recovery, ischemic lesion diminished while microvasculature regeneration progressed. Furthermore, the newly formed capillary network surrounding the lesion had a highly organized and directional structure with a large percentage of capillary segments stretching towards the ischemic center. Deeper de novo microvasculature had a better organized structure than shallower one, suggesting that deeper tissue oxygenation has a higher dependence on capillary network. Our stalling event analysis revealed that ischemic stroke increased both stalling density and incidence. As stalling can lead to cerebral blood flow reduction, targeting post-stroke stalling could be useful to develop new treatment for stroke

Development of a diagnostic sensor for measuring blood cell concentrations during haemoconcentration

Background: HemoSep® is a commercial ultrafiltration and haemoconcentration device for the concentration of residual bypass blood following surgery. This technology is capable of reducing blood loss in cardiac and other types of "clean site" procedures, including paediatric surgery. Clinical feedback suggested that the device would be enhanced by including a sensor technology capable of discerning the concentration level of the processed blood product. We sought to develop a novel sensor that can, using light absorption, give an accurate estimate of packed cell volume (PCV). Materials and methods: A sensor-housing unit was 3D printed and the factors influencing the sensor's effectiveness – supply voltage, sensitivity and emitter intensity - were optimised. We developed a smart system, using comparator circuitry capable of visually informing the user when adequate PCV levels (⩾35%) are attained by HemoSep® blood processing, which ultimately indicates that the blood is ready for autotransfusion. Results: Our data demonstrated that the device was capable of identifying blood concentration at and beyond the 35% PCV level. The device was found to be 100% accurate at identifying concentration levels of 35% from a starting level of 20%. Discussion: The sensory capability was integrated into HemoSep's® current device and is designed to enhance the user’s clinical experience and to optimise the benefits of HemoSep® therapy. The present study focused on laboratory studies using bovine blood. Further studies are now planned in the clinical setting to confirm the efficacy of the device

On the Indeterminates of Glaucoma:the Controversy of Arterial Blood Pressure and Retinal Perfusion

Glaucoma is a chronic eye disease characterized by thinning of the retina, death of ganglion cells, and progressive loss of vision, eventually leading to blindness. The prevalence of glaucoma is estimated at 1-3% of those over 40 years old. With a constantly aging population, this number is expected to increase significantly over the next 10 years. Even with treatment, about 15% of people with glaucoma currently develop residual vision or tunnel vision and eventually become blind or partially sighted. The mechanisms behind ganglion cell death are poorly understood. Elevated eye pressure is the main risk factor for glaucoma, but treatment in the form of medication, laser, or surgery can only slow the decline, not stop it. In addition, high intraocular pressure is neither necessary nor sufficient for the development of glaucoma, indicating the existence of other unknown risk factors. It has been established that the death of ganglion cells results in a decreased oxygen demand and a concomitant decrease in blood flow. However, there is also a hypothesis that reduced or unstable blood supply is not only a consequence, but also a cause of glaucoma. This is known as the ‘chicken-egg’ dilemma in glaucoma. It is supported by the observation that the risk of developing glaucoma is higher in people with very low blood pressure (sometimes even as a result of overtreatment of high blood pressure).This dissertation is an attempt to methodically examine whether blood pressure can be linked to changes in the retina that could suggest susceptibility to glaucoma. For this purpose, we analyze epidemiological data from the Groningen Longitudinal Glaucoma Study, we use advanced imaging techniques to model the microcirculation, and we describe its relationship with the neural structure and oxygen consumption of the retina. We provide evidence leaning towards the existence of a vascular component, likely pertinent to glaucoma

Measurement and clinical implications of choroidal thickness in patients with inflammatory bowel disease

ABSTRACTPurpose:Ocular inflammation is a frequent extraintestinal manifestation of inflammatory bowel disease (IBD) and may parallel disease activity. In this study, we evaluated the utility of a choroidal thickness measurement in assessing IBD activity.Methods:A total of 62 eyes of 31 patients with IBD [Crohn's disease (CD), n=10 and ulcerative colitis (UC), n=21] and 104 eyes of 52 healthy blood donors were included in this study. Choroidal thickness was measured using enhanced depth imaging optical coherence tomography. The Crohn's disease activity index (CDAI) and the modified Truelove Witts score were used to assess disease activity in CD and UC, respectively.Results:No significant differences in mean subfoveal, nasal 3000 μm, or temporal 3000 μm choroidal thickness measurements (P>0.05 for all) were observed between IBD patients and healthy controls. Age, smoking, CD site of involvement (ileal and ileocolonic involvement), CDAI, CD activity, and UC endoscopic activity index were all found to be significantly correlated with choroidal thickness by univariate analysis (P<0.05). Smoking (P<0.05) and the CD site of involvement (P<0.01) were the only independent parameters associated with increased choroidal thickness at all measurement locations.Conclusions:Choroidal thickness is not a useful marker of disease activity in patients with IBD but may be an indicator of ileal involvement in patients with CD

Intact in vivo visualization of telencephalic microvasculature in medaka using optical coherence tomography

To date, various human disease models in small fish—such as medaka (Oryzias lapties)--have been developed for medical and pharmacological studies. Although genetic and environmental homogeneities exist, disease progressions can show large individual differences in animal models. In this study, we established an intact in vivo angiographic approach and explored vascular networks in the telencephalon of wild-type adult medaka using the spectral-domain optical coherence tomography. Our approach, which required neither surgical operations nor labeling agents, allowed to visualize blood vessels in medaka telencephala as small as about 8 µm, that is, almost the size of the blood cells of medaka. Besides, we could show the three-dimensional microvascular distribution in the medaka telencephalon. Therefore, the intact in vivo imaging via optical coherence tomography can be used to perform follow-up studies on cerebrovascular alterations in metabolic syndrome and their associations with neurodegenerative disease models in medaka