Multimodal CARS microscopy determination of the impact of diet on macrophage infiltration and lipid accumulation on plaque formation in ApoE-deficient mice

We characterized several cellular and structural features of early stage Type II/III atherosclerotic plaques in an established model of atherosclerosis—the ApoE-deficient mouse—by using a multimodal, coregistered imaging system that integrates three nonlinear optical microscopy (NLOM) contrast mechanisms: coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), and two-photon excitation fluorescence (TPEF). Specifically, the infiltration of lipid-rich macrophages and the structural organization of collagen and elastin fibers were visualized by CARS, SHG, and TPEF, respectively, in thick tissue specimens without the use of exogenous labels or dyes. Label-free CARS imaging of macrophage accumulation was confirmed by histopathology using CD68 staining. A high-fat, high-cholesterol Western diet resulted in an approximate 2-fold increase in intimal plaque area, defined by CARS signals of lipid-rich macrophages. Additionally, analysis of collagen distribution within lipid-rich plaque regions revealed nearly a 4-fold decrease in the Western diet–fed mice, suggesting NLOM sensitivity to increased matrix metalloproteinase (MMP) activity and decreased smooth muscle cell (SMC) accumulation. These imaging results provide significant insight into the structure and composition of early stage Type II/III plaque during formation and allow for quantitative measurements of the impact of diet and other factors on critical plaque and arterial wall features.     

Scar tissue classification using nonlinear optical microscopy and discriminant analysis

This paper addresses the scar tissue maturation process that occurs stepwise, and calls for reliable classification. The structure of collagen imaged by nonlinear optical microscopy (NLOM) in post‐burn hypertrophic and mature scar, as well as in normal skin, appeared to distinguish these maturation steps. However, it was a discrimination analysis, demonstrated here, that automated and quantified the scar tissue maturation process. The achieved scar classification accuracy was as high as 96%. The combination of NLOM and discrimination analysis is believed to be instrumental in gaining insight into the scar formation, for express diagnosis of scar and surgery planning. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Molecular histology of arteries: mass spectrometry imaging as a novel ex vivo tool to investigate atherosclerosis

Atherosclerosis is usually the underlying cause of a fatal event such as myocardial infarction or ictus. The atherome plaque develops silently and asymptomatically within the arterial intima layer. In this context, the possibility to analyze the molecular content of arterial tissue while preserving each molecule’s specific localization is of great interest as it may reveal further insights into the physiopathological changes taking place. Mass spectrometry imaging (MSI) enables the spatially resolved molecular analysis of proteins, peptides, metabolites, lipids and drugs directly in tissue, with a resolution sufficient to reveal molecular features specific to distinct arterial structures. MSI represents a novel ex vivo imaging tool still underexplored in cardiovascular diseases. This review focuses on the MSI technique applied to cardiovascular disease and covers the main contributions to date, ongoing efforts, the main challenges and current limitations of MSI.     

The use of small angle light scattering in assessing strain induced collagen degradation in arterial tissue ex vivo

Collagen is the predominant load bearing component in many soft tissues including arterial tissue and is therefore critical in determining the mechanical integrity of such tissues. Degradation of collagen fibres is hypothesized to be a strain dependent process whereby the rate of degradation is affected by the magnitude of strain applied to the collagen fibres. The aim of this study is to investigate the ability of small angle light scattering (SALS) imaging to identify strain dependent degradation of collagen fibres in arterial tissue ex vivo, and determine whether a strain induced protection mechanism exists in arterial tissue as observed in pure collagen and other collagenous tissues. SALS was used in combination with histological and second harmonic generation (SHG) analysis to determine the collagen fibre architecture in arterial tissue subjected to strain directed degradation. SALS alignment analysis identified statistically significant differences in fibre alignment depending on the strain magnitude applied to the tissue. These results were also observed using histology and SHG. Our findings suggest a strain protection mechanism may exist for arterial collagen at intermediate strain magnitudes between 0% and 25%. These findings may have implications for the onset and progression of arterial disease where changes in the mechanical environment of arterial tissue may lead to changes in the collagen degradation rate.     

Application of Numerical Methods to Elasticity Imaging

Elasticity imaging can be understood as the intersection of the study of biomechanical properties, imaging sciences, and physics. It was mainly motivated by the fact that pathological tissue presents an increased stiffness when compared to surrounding normal tissue. In the last two decades, research on elasticity imaging has been an international and interdisciplinary pursuit aiming to map the viscoelastic properties of tissue in order to provide clinically useful information. As a result, several modalities of elasticity imaging, mostly based on ultrasound but also on magnetic resonance imaging and optical coherence tomography, have been proposed and applied to a number of clinical applications: cancer diagnosis (prostate, breast, liver), hepatic cirrhosis, renal disease, thyroiditis, arterial plaque evaluation, wall stiffness in arteries, evaluation of thrombosis in veins, and many others. In this context, numerical methods are applied to solve forward and inverse problems implicit in the algorithms in order to estimate viscoelastic linear and nonlinear parameters, especially for quantitative elasticity imaging modalities. In this work, an introduction to elasticity imaging modalities is presented. The working principle of qualitative modalities (sonoelasticity, strain elastography, acoustic radiation force impulse) and quantitative modalities (Crawling Waves Sonoelastography, Spatially Modulated Ultrasound Radiation Force (SMURF), Supersonic Imaging) will be explained. Subsequently, the areas in which numerical methods can be applied to elasticity imaging are highlighted and discussed. Finally, we present a detailed example of applying total variation and AM-FM techniques to the estimation of elasticity.     

Intra-arterial catheter for simultaneous microstructural and molecular imaging in vivo

Advancing understanding of human coronary artery disease requires new methods that can be used in patients for studying atherosclerotic plaque microstructure in relation to the molecular mechanisms that underlie its initiation, progression and clinical complications, including myocardial infarction and sudden cardiac death. Here we report a dual-modality intra-arterial catheter for simultaneous microstructural and molecular imaging in vivo using a combination of optical frequency domain imaging (OFDI) and near-infrared fluorescence (NIRF) imaging. By providing simultaneous molecular information in the context of the surrounding tissue microstructure, this new catheter could provide new opportunities for investigating coronary atherosclerosis and stent healing and for identifying high-risk biological and structural coronary arterial plaques in vivo.     

Thematic review series: patient-oriented research. Imaging atherosclerosis: state of the art

The ability to image obstructive arterial disease brought about a revolution in clinical cardiovascular care; the development of newer technologies that image arterial wall thicknesses, areas, volumes, and composition allows valid imaging of atherosclerosis for the first time. Development of noninvasive imaging of atherosclerosis has further led to a quantum shift in research in the field by enabling the study of asymptomatic populations and thus allowing investigators to focus on preclinical disease without the many biases associated with the study of symptomatic patients. These noninvasive investigations have broad implications for clinical care as well. Coronary angiography, computed tomographic (CT) imaging of coronary calcium, intravascular ultrasound, multidetector CT angiography, B mode ultrasound of the carotid arteries, and MRI of the carotid arteries all have unique strengths and weaknesses for imaging atherosclerosis. Certain of these techniques are extremely useful as outcome variables for clinical trials, and others are uniquely useful as predictors of the risk of cardiovascular disease. All are informative in one way or another with regard to the role of plaque remodeling and composition in disease causation. CT and MRI technology are advancing very rapidly, and research and clinical uses of these imaging modalities promise to further advance our understanding of atherosclerosis and its prevention.     

Imaging angiogenesis

Angiogenesis represents the formation of new capillaries by cellular outgrowth from existing microvessels and plays a critical role in the response to ischemia associated with peripheral arterial disease and myocardial infarction. Imaging of angiogenesis would be valuable in risk stratification of patients with arterial occlusive disease. The progress in noninvasive imaging strategies to assess angiogenesis has been made possible with the availability of many technological advances, which include dedicated hybrid SPECT-CT and PET-CT systems and agents targeted at molecular markers of the angiogenic process, involving both receptor-probe interactions and reporter gene technology. These novel targeted approaches for imaging angiogenesis will complement standard imaging of physiological parameters and will play a crucial role for evaluation of therapeutic interventions to promote angiogenesis.     

An automatic diagnostic system of coronary artery lesions in Kawasaki disease using intravascular optical coherence tomography imaging

Intravascular optical coherence tomography (IV‐OCT) is a light‐based imaging modality with high resolution, which employs near‐infrared light to provide tomographic intracoronary images. Morbidity caused by coronary heart disease is a substantial cause of acute coronary syndrome and sudden cardiac death. The most common intracoronay complications caused by coronary artery disease are intimal hyperplasia, calcification, fibrosis, neovascularization and macrophage accumulation, which require efficient prevention strategies. OCT can provide discriminative information of the intracoronary tissues, which can be used to train a robust fully automatic tissue characterization model based on deep learning. In this study, we aimed to design a diagnostic model of coronary artery lesions. Particularly, we trained a random forest using convolutional neural network features to distinguish between normal and diseased arterial wall structure. Then, based on the arterial wall structure, fully convolutional network is designed to extract the tissue layers in normal cases, and pathological tissues regardless of lesion type in pathological cases. Then, the type of the lesions can be characterized with high precision using our previous model. The results demonstrate the robustness of the model with the approximate overall accuracy up to 90%.      

The Acid-Base Status in Cyanotic Congenital Heart Disease

The relation between oxygen consumption, metabolic status and prognosis was studied in two infants with identically low arterial oxygen tensions (20 mm. Hg) due to cyanotic congenital heart disease. The first patient had low oxygen consumption, arterial blood acidosis and increased arterial lactate, and died at the age of 36 hours. The second had normal oxygen consumption, arterial acid-base balance, lactate and pyruvate, and survived. Since arterial oxygen tensions were similar in both, it was concluded that compensatory factors, such as cardiac output, pulmonary and systemic blood flow and increased oxygen saturation at normal pH levels (Bohr effect), are important in maintaining tissue oxidation and preventing anaerobiosis and lactate production. The importance of a knowledge of acid-base status in the immediate prognosis of cyanotic congenital heart disease is stressed. The treatment of acidosis by buffer therapy may be an important palliative, improving cardiac output and tissue oxidation, and should be undertaken as soon as possible before irreversible cellular damage has occurred.     

Characterization of vascular strain during in-vitro angioplasty with high-resolution ultrasound speckle tracking

Background

Ultrasound elasticity imaging provides biomechanical and elastic properties of vascular tissue, with the potential to distinguish between tissue motion and tissue strain. To validate the ability of ultrasound elasticity imaging to predict structurally defined physical changes in tissue, strain measurement patterns during angioplasty in four bovine carotid artery pathology samples were compared to the measured physical characteristics of the tissue specimens.

Methods

Using computational image-processing techniques, the circumferences of each bovine artery specimen were obtained from ultrasound and pathologic data.

Results

Ultrasound-strain-based and pathology-based arterial circumference measurements were correlated with an R² value of 0.94 (p = 0.03). The experimental elasticity imaging results confirmed the onset of deformation of an angioplasty procedure by indicating a consistent inflection point where vessel fibers were fully unfolded and vessel wall strain initiated.

Conclusion

These results validate the ability of ultrasound elasticity imaging to measure localized mechanical changes in vascular tissue.     

Multiples défauts segmentaires et sous-segmentaires de perfusion discordants avec une angiographie pulmonaire normale et une forte suspicion de maladie veino-occlusive pulmonaire

We report the case of a 55-year-old woman presenting stage-3-4 dyspnea over the last 5 years with no other clinical symptoms. Pulmonary arterial hypertension (PAH) was evidenced by echocardiography. However, there was no argument for chronic thromboembolic disease during pulmonary angiography. The pulmonary scintigraphy showed homogeneous ventilation on both lung fields, with multiple bilateral segmental and subsegmental perfusion defects, however. These perfusion abnormalities with a normal pulmonary angiography led to the diagnosis of pulmonary veno-occlusive disease (PVOD) taking into account the clinical context, imaging findings, and laboratory and functional assessments. PVOD is a clinicopathological entity characterized by the occlusion or narrowing of the pulmonary veins and venules by fibrous tissue, leading to clinical manifestations that are, in many ways similar to PAH. PVOD remains poorly understood and it is difficult to diagnose and treat. However, PVOD needs to be known by clinicians and considered in the differential diagnosis of PAH.     

Oxidized low-density lipoprotein increases cultured human endothelial cell tissue factor activity and reduces protein C activation

Increasing evidence suggests that the formation of oxidized low-density lipoprotein (Ox-LDL) in vivo is associated with the development of atherosclerotic vascular disease. We investigated the effects of Ox-LDL on two vascular endothelial cell coagulant properties, tissue factor expression, and protein C activation. The Ox-LDL increased human arterial and venous endothelial cell tissue factor activity, with 100 micrograms/ml of Ox-LDL increasing factor activity fourfold. Native LDL modified by incubation with cultured human arterial and venous endothelial cells also induced endothelial cell tissue factor activity. This modification was blocked by coincubation with the antioxidants, probucol or ascorbic acid. It was determined, based on inhibition by known scavenger receptor antagonists (fucoidin, dextran sulfate), that binding of Ox-LDL via the acetyl LDL (scavenger) receptor was partially responsible for the increase in tissue factor expression. Whereas endothelial cell tissue factor expression was increased by incubation with Ox-LDL, protein C activation was reduced approximately 80% by incubating cultured endothelial cells with Ox-LDL. The effect of Ox-LDL on protein C activation was not inhibited by antagonists to the scavenger receptor. These data indicating that an atherogenic lipoprotein can regulate key vascular coagulant activities provide an additional link between vascular disease and thrombosis.     

Quantitative Amyloid Imaging Using Image-Derived Arterial Input Function

Amyloid PET imaging is an indispensable tool widely used in the investigation, diagnosis and monitoring of Alzheimer’s disease (AD). Currently, a reference region based approach is used as the mainstream quantification technique for amyloid imaging. This approach assumes the reference region is amyloid free and has the same tracer influx and washout kinetics as the regions of interest. However, this assumption may not always be valid. The goal of this work is to evaluate an amyloid imaging quantification technique that uses arterial region of interest as the reference to avoid potential bias caused by specific binding in the reference region. 21 participants, age 58 and up, underwent Pittsburgh compound B (PiB) PET imaging and MR imaging including a time-of-flight (TOF) MR angiography (MRA) scan and a structural scan. FreeSurfer based regional analysis was performed to quantify PiB PET data. Arterial input function was estimated based on coregistered TOF MRA using a modeling based technique. Regional distribution volume (V_T) was calculated using Logan graphical analysis with estimated arterial input function. Kinetic modeling was also performed using the estimated arterial input function as a way to evaluate PiB binding (DVR_kinetic) without a reference region. As a comparison, Logan graphical analysis was also performed with cerebellar cortex as reference to obtain DVR_REF. Excellent agreement was observed between the two distribution volume ratio measurements (r>0.89, ICC>0.80). The estimated cerebellum V_T was in line with literature reported values and the variability of cerebellum V_T in the control group was comparable to reported variability using arterial sampling data. This study suggests that image-based arterial input function is a viable approach to quantify amyloid imaging data, without the need of arterial sampling or a reference region. This technique can be a valuable tool for amyloid imaging, particularly in population where reference normalization may not be accurate.     

Capacities of computed tomographic angiography in the diagnosis and estimation of the site, pattern, and degree of vascular bed lesion in nonspecific aortoarteritis

Nonspecific aortoarteritis (NAA) is a chronic inflammatory disease that affects the aorta and its branches. The clinical manifestations of this disease are of a great variety and depend on the site of a lesion and the stage of the disease. A wide range of highly informative noninvasive imaging techniques, such as duplex scanning (DS), magnetic resonance imaging (MRI), and computed tomography (CT) of the aorta and its branches, are used to make a more accurate diagnosis and to determine the site and extent of a vascular bed lesion. The given clinical example suggests that CT angiography of the aorta and its branches is a high-precision technique in determining the site of vascular bed lesion in patients with NAA and the pattern and extent of arterial involvement and that it may be used for both the diagnosis of the disease at its developmental stages and the monitoring of the vessels during pathogenetic therapy.     

Real-time ultrasonic imaging of the peripheral arteries: Technique, normal anatomy, and pathology

Real-time ultrasonic echo imaging of peripheral arteries promises to facilitate the management of selected patients with peripheral and extracranial arterial disease. This report outlines the technique of imaging the carotid system and portions of the arteries that supply the lower extremities. It also discusses the normal and pathologic anatomy of these arteries.     

Molecular imaging of autoimmune diseases and inflammation

Molecular imaging methods allow the noninvasive detection and localization of specific molecules. Agents that report on molecular disease biomarkers can be used to diagnose and monitor disease. Many inflammatory diseases have molecular signatures within altered tissues. Although tissue biopsy is still the gold standard for detecting these signatures, several molecular imaging markers have been developed. Pharmacologic agents that block specific immune molecules have recently entered the clinic, and these drugs have already transformed the way we care for patients with immune-mediated diseases. The use of immunomodulatory drugs is usually guided by clinical assessment of the patient's response. Unfortunately, clinical assessment may miss the signs of inflammation, and many of the serologic markers of immune-mediated diseases correlate poorly with the underlying inflammatory activity within target tissues. Molecular imaging methods have the potential to improve our ability to detect and characterize tissue inflammation. We discuss some of the molecular signatures of immune activation and review molecular imaging methods that have been developed to detect active tissue inflammation.     

Molecular-genetic imaging: a nuclear medicine-based perspective

Molecular imaging is a relatively new discipline, which developed over the past decade, initially driven by in situ reporter imaging technology. Noninvasive in vivo molecular-genetic imaging developed more recently and is based on nuclear (positron emission tomography [PET], gamma camera, autoradiography) imaging as well as magnetic resonance (MR) and in vivo optical imaging. Molecular-genetic imaging has its roots in both molecular biology and cell biology, as well as in new imaging technologies. The focus of this presentation will be nuclear-based molecular-genetic imaging, but it will comment on the value and utility of combining different imaging modalities. Nuclear-based molecular imaging can be viewed in terms of three different imaging strategies: (1) "indirect" reporter gene imaging; (2) "direct" imaging of endogenous molecules; or (3) "surrogate" or "bio-marker" imaging. Examples of each imaging strategy will be presented and discussed. The rapid growth of in vivo molecular imaging is due to the established base of in vivo imaging technologies, the established programs in molecular and cell biology, and the convergence of these disciplines. The development of versatile and sensitive assays that do not require tissue samples will be of considerable value for monitoring molecular-genetic and cellular processes in animal models of human disease, as well as for studies in human subjects in the future. Noninvasive imaging of molecular-genetic and cellular processes will complement established ex vivo molecular-biological assays that require tissue sampling, and will provide a spatial as well as a temporal dimension to our understanding of various diseases and disease processes.     

Prediction of fibre architecture and adaptation in diseased carotid bifurcations

Many studies have used patient-specific finite element models to estimate the stress environment in atherosclerotic plaques, attempting to correlate the magnitude of stress to plaque vulnerability. In complex geometries, few studies have incorporated the anisotropic material response of arterial tissue. This paper presents a fibre remodelling algorithm to predict the fibre architecture, and thus anisotropic material response in four patient-specific models of the carotid bifurcation. The change in fibre architecture during disease progression and its affect on the stress environment in the plaque were predicted. The mean fibre directions were assumed to lie at an angle between the two positive principal strain directions. The angle and the degree of dispersion were assumed to depend on the ratio of principal strain values. Results were compared with experimental observations and other numerical studies. In non-branching regions of each model, the typical double helix arterial fibre pattern was predicted while at the bifurcation and in regions of plaque burden, more complex fibre architectures were found. The predicted change in fibre architecture in the arterial tissue during plaque progression was found to alter the stress environment in the plaque. This suggests that the specimen-specific anisotropic response of the tissue should be taken into account to accurately predict stresses in the plaque. Since determination of the fibre architecture in vivo is a difficult task, the system presented here provides a useful method of estimating the fibre architecture in complex arterial geometries.     

Quantitative models of the rat pulmonary arterial tree morphometry applied to hypoxia-induced arterial remodeling.

Little is known about the constituent hemodynamic consequences of structural changes that occur in the pulmonary arteries during the onset and progression of pulmonary arterial remodeling. Many disease processes are known to be responsible for vascular remodeling that leads to pulmonary arterial hypertension, cor pulmonale, and death. Histology has been the primary tool for evaluating pulmonary remodeling, but it does not provide information on intact vascular structure or the vessel mechanical properties. This study is an extension of our previous work in which we developed an alternative imaging technique to evaluate pulmonary arterial structure. The lungs from Sprague-Dawley rats were removed, perfusion analysis was performed on the isolated lungs, and then an X-ray contrast agent was used to fill the arterial network for imaging. The lungs were scanned over a range of intravascular pressures by volumetric micro-computed tomography, and the arterial morphometry was mapped and measured in the reconstructed isotropic volumes. A quantitative assessment of hemodynamic, structural, and biomechanical differences between rats exposed for 21 days to hypoxia (10% O(2)) or normoxia (21.0% O(2)) was performed. One metric, the normalized distensibility of the arteries, is significantly (P < 0.001) larger [0.025 +/- 0.0011 (SE) mmHg(-1)] (n = 9) in normoxic rats compared with hypoxic [0.015 +/- 0.00077 (SE) mmHg(-1)] (n = 9). The results of the study show that these models can be applied to the Sprague-Dawley rat data and, specifically, can be used to differentiate between the hypoxic and the control groups.