Human Coronary Arterial Wall Research Articles

Recent Advances in Fluorescent Angioscopy for Molecular Imaging of Human Atherosclerotic Coronary Plaque.

Purpose of Review: In vivo imaging of the native substances, including lipoproteins, that comprise human atherosclerotic plaques is currently beyond the scope of any available imaging techniques. Color and near-infrared fluorescent angioscopy (CFA and NIRFA, respectively) systems have been recently developed for molecular imaging of lipoproteins within the human coronary arterial wall ex vivo and/or in vivo. The author reviews recent findings on lipoprotein deposition in human coronary plaques obtained by these imaging techniques.Recent Findings: Using specific biomarkers, native pro-atherogenic substances such as oxidized low-density lipoprotein (ox-LDL), LDL, triglycerides (TG), apolipoprotein B-100 (ApoB-100), and lysophosphatidylcholine (LPC), and the anti-atherogenic substance such as high-density lipoprotein (HDL) were visualized by CFA, and LDL and cholesterol by NIRFA, in coronary plaques obtained from autopsy subjects. The relationship between incidence and plaque morphology differed for each substance. The incidence of ox-LDL and LDL on color fluorescence microscopy correlated well with that observed using immunohistochemical techniques. During coronary catheterization in patients, ox-LDL, LDL, and HDL in coronary plaques were visualized by CFA or NIRFA.Conclusions: Using CFA or NIRFA, the distribution of the major native pro-atherogenic and antiatherogenic lipoproteins and their components within human coronary plaques can be evaluated ex vivo and/or in vivo. Fluorescent angioscopy could help our understanding of the molecular mechanisms of coronary atherosclerosis and in the evaluation of the effects of therapy targeting the substances comprising atherosclerotic coronary plaques.

Molecular Imaging of Native Low-Density Lipoprotein by Near-Infrared Fluorescent Angioscopy in Human Coronary Plaques

Low-density lipoprotein (LDL) is an important risk factor for coronary artery disease, but its localization within the human coronary arterial wall is poorly understood. Imaging of LDL in 30 coronary arteries excised from 15 subjects who underwent autopsy was performed using near-infrared fluorescent angioscopy system and using indocyanine green dye as a biomarker of LDL. The percentage incidence of LDL in 28 normal segments, 24 white plaques (early stage of plaque growth), and 21 yellow plaques (mature stage of plaque) classified by conventional angioscopy, was 14.2, 79.1 (p <0.01 vs normal segments and p <0.05 vs yellow plaques), and 28.5, respectively. Coronary near-infrared fluorescent angioscopy showed similar results in 7 patients in vivo. Our results suggested that LDL begins to deposit in the human coronary arterial wall in the early stage of atherosclerosis, increasingly deposits with plaque growth and decreases in the mature stage; and therefore, molecular therapy targeting LDL should be started before plaque maturation.

Diagnostic fiber-based optical imaging catheters

Fiber optics miniaturization and advances in signal acquisition and processing have allowed the development of fiber-based optical imaging catheters that permit instantaneous luminal organ imaging. This technique is applicable in clinical settings for diagnosing various diseases. Intravascular optical coherence tomography (IV-OCT), a catheter-based optical imaging technique, acquires high-resolution cross-sectional human coronary arterial wall imaging, enabling precise assessment of coronary atherosclerosis. OCT with a ballooncentering catheter or a tethered capsule acquires comprehensive three-dimensional images of the distal esophagus for diagnostic imaging in patients with esophageal diseases, including Barrett’s esophagus. Spectrally encoded confocal microscopy (SECM), an advanced type of confocal microscopy that uses diffraction grating and a broadband laser source to laterally scan the sample without mechanical motion, has been developed as a tethered confocal endomicroscopy capsule to diagnose and monitor eosinophilic esophagitis, an allergic condition in the esophageal wall. In this review, the authors describe the recent development of fiber-based imaging catheters with rotary scanning for diagnosing various diseases in luminal organs, including the coronary artery and esophagus. Further developments, including miniaturization of optics, increased speed, and multimodal acquisition, could significantly improve diagnostic capability to improve patient care.

Imaging of native high-density lipoprotein in human coronary plaques by color fluorescent angioscopy.

High-density lipoprotein (HDL) plays a key role in reverse cholesterol transport, and halts the progression of atherosclerosis. The aim of the present study was to visualize native HDL in the human coronary arterial wall. The fluorescence characteristics of HDL were investigated by color fluorescent microscopy (CFM) using excitation at 470 nm and emission at 515 nm with Fast green dye (FG) as the biomarker. HDL in 30 normal coronary segments, and in 25 white and 25 yellow plaques in excised human coronary arteries, was visualized by color fluorescent angioscopy (CFA) and CFM. Localization of HDL visualized by CFM was compared with that stained by immunostaining using an anti-HDL antibody. FG elicited a characteristic brown fluorescence of HDL. By CFA, the percent incidence of HDL in normal segments, white (early stage of plaque growth) and yellow (advanced stage of plaque growth) plaques was, respectively, 33%, 76% (P<0.05 vs. normal segments and yellow plaques) and 21%. Localization of HDL visualized by CFM did not differ from that stained by immunostaining. In the human coronary arterial wall, HDL deposits infrequently in normal segments, but increasingly deposits with plaque formation, and decreases in the advanced stage of plaque growth.

Localization of oxidized low-density lipoprotein and its relation to plaque morphology in human coronary artery.

ObjectivesOxidized low-density lipoprotein (oxLDL) plays a key role in the formation of atherosclerotic plaques. However, its localization in human coronary arterial wall is not well understood. The present study was performed to visualize deposition sites and patterns of native oxLDL and their relation to plaque morphology in human coronary artery.MethodsEvans blue dye (EB) elicits a violet fluorescence by excitation at 345-nm and emission at 420-nm, and a reddish-brown fluorescence by excitation at 470-nm and emission at 515-nm characteristic of oxLDL only. Therefore, native oxLDL in excised human coronary artery were investigated by color fluorescent microscopy (CFM) using EB as a biomarker.Results(1) By luminal surface scan with CFM, the % incidence of oxLDL in 38 normal segments, 41 white plaques and 32 yellow plaques that were classified by conventional angioscopy, was respectively 26, 44 and 94, indicating significantly (p<0.05) higher incidence in the latter than the former two groups. Distribution pattern was classified as patchy, diffuse and web-like. Web-like pattern was observed only in yellow plaques with necrotic core. (2) By transected surface scan, oxLDL deposited within superficial layer in normal segments and diffusely within both superficial and deep layers in white and yellow plaques. In yellow plaques with necrotic core, oxLDL deposited not only in the marginal zone of the necrotic core but also in the fibrous cap.ConclusionTaken into consideration of the well-known process of coronary plaque growth, the results suggest that oxLDL begins to deposit in human coronary artery wall before plaque formation and increasingly deposits with plaque growth, exhibiting different deposition sites and patterns depending on morphological changes.

Molecular Imaging of Apolipoprotein B-100 in Human Coronary Plaques by Color Fluorescent Angioscopy and Microscopy

Apolipoprotein B-100 (ApoB-100) is an important risk factor for coronary artery disease. However, its localization in human coronary plaques is not well understood. The present study was performed to visualize ApoB-100 in human coronary artery wall. Deposition of native ApoB-100 in excised human coronary plaques and normal segments classified by conventional angioscopy was investigated by color fluorescent angioscopy (CFA) and microscopy (CFM) using Nile blue dye (NB) which elicits a golden fluorescence characteristic of ApoB-100 as a biomarker. By CFA, the % incidence of ApoB-100 was 20 in 40 normal segments, 38 in 42 white, and 11 in 35 yellow plaques (P < 0.05 versus white plaques). There was no significant difference in detection sensitivity between CFA and luminal surface scan by CFM. By CFM transected surface scan, ApoB-100 deposited in superficial, deep, and/or in both layers. Deposition in both layers was frequently observed in white plaques and yellow plaques without necrotic core (NC), less frequently in normal segments, and rarely in yellow plaques with NC. (1) Taking into consideration the well known process of plaque growth, the results suggest that ApoB-100 begins to deposit before plaque formation, increasingly deposits with plaque growth, and disappears after necrotic core formation. (2) CFA is feasible for imaging of ApoB-100 in human coronary artery wall.

Localization of Native High-Density Lipoprotein and Its Relation to Plaque Morphology in Human Coronary Artery

High-density lipoprotein (HDL) plays a key role in reverse cholesterol transport, and halts the progression of atherosclerosis. However, its localization in human vascular wall is not well understood. We discovered that by exciting at 470-nm and emitting at 515-nm light wavelengths, Fast green dye (FG) elicits brown fluorescence characteristic of HDL only. Therefore, the localization of native HDL in normal segments and plaques in excised human coronary artery was investigated by scanning their transected surface with color fluorescent microscopy (CFM) using FG as a biomarker, and the relationships between the localization of HDL and morphology of plaques and normal segments classified by conventional angioscopy and histology were examined. The % incidence of HDL in 13 normal segments (NS) with thin (≤ 200 µm) intima, 28 NS with thick (200 µm <) intima, 41 white plaques (early stage of plaque growth), 15 yellow plaques (Y) without necrotic core (NC), and 20 Y with NC (advanced stage of plaque growth), was 30, 71 (P < 0.05 versus NS with thin intima and Y with NC), 83 (P < 0.05 versus NS with thin intima and Y with NC), 60, and 35, respectively. HDL begins to deposit in human coronary arterial wall in the early stage of atherosclerosis and deposits increase with plaque growth, but HDL decreases in plaques at an advanced stage of growth.

Molecular Imaging of Low-density Lipoprotein in Human Coronary Plaques by Color Fluorescent Angioscopy and Microscopy

ObjectivesLow-density lipoprotein (LDL) is an important risk factor for coronary artery disease. However, its localization in human coronary plaques is not well understood. The present study was performed to visualize LDL in human coronary artery wall.Methods(1) The fluorescence characteristic of LDL was investigated by color fluorescent microscopy (CFM) with excitation at 470-nm and emission at 515-nm using Nile blue dye (NB) as a biomarker. (2) Native LDL in 40 normal segments, 42 white plaques and 35 yellow plaques (20 with necrotic core) of human coronary arteries was investigated by color fluorescent angioscopy (CFA) and CFM.Results(1) NB elicited a brown, golden and red fluorescence characteristic of LDL, apolipoprotein B-100, and lysophosphatidylcholine/triglyceride, respectively. (2) The % incidence of LDL in normal segments, white, and yellow plaques was 25, 38 and 14 by CFA and 42, 42 and 14 by CFM scan of their luminal surface, respectively, indicating lower incidence (p<0.05) of LDL in yellow plaques than white plaques, and no significant differences in detection sensitivity between CFA and CFM. By CFM transected surface scan, LDL deposited more frequently and more diffusely in white plaques and yellow plaques without necrotic core (NC) than normal segments and yellow plaques with NC. LDL was localized to fibrous cap in yellow plaques with NC. Co-deposition of LDL with other lipid components was observed frequently in white plaques and yellow plaques without NC.Conclusions(1) Taken into consideration of the well-known process of coronary plaque growth, the results of the present study suggest that LDL begins to deposit before plaque formation; increasingly deposits with plaque growth, often co-depositing with other lipid components; and disappears after necrotic core formation. (2) CFA is feasible for visualization of LDL in human coronary artery wall.

Musashi-1 expression in atherosclerotic arteries and its relevance to the origin of arterial smooth muscle cells: Histopathological findings and speculations

The origin of smooth muscle cells in developing atherosclerotic lesions is a controversial topic with accumulating evidence indicating that at least some arterial smooth muscle cells might originate from bone marrow-derived smooth muscle cell precursors circulating in the blood. The stem cell markers currently used for the identification of stem cells in the arterial intima can be expressed by a number of different cell types residing in the arterial wall, such as mast cells, endothelial cells and dendritic cells, which can make interpretation of the data obtained somewhat ambiguous. In the present study we examined whether the putative intestinal stem cell marker Musashi-1 is expressed in the arterial wall. Using a multiplexed tandem polymerase chain reaction method (MT-PCR) and immunohistochemistry, Musashi-1 expression was revealed in human coronary arterial wall tissue segments, and this finding was followed by the demonstration of significantly higher expression levels of Musashi-1 in atherosclerotic plaques compared with those in undiseased intimal sites. Double immunohistochemistry demonstrated that in the arterial wall Musashi-1 positive cells either did not display any specific markers of cells that are known to reside in the arterial intima or Musashi-1 was co-expressed by smooth muscle α-actin positive cells. Some Musashi-1 positive cells were found along the luminal surface of arteries as well as within microvessels formed in atherosclerotic plaques by neovascularization, which supports the possibility that Musashi-1 positive cells might intrude into the arterial wall from the blood and might even represent circulating smooth muscle cell precursors.

Age and gender effects on apoptosis in the human coronary arterial wall

The relationship between aging and apoptosis remains unclear. We wondered whether apoptosis could be enhanced in arterial aging in the absence of overt or advanced arterial disease. Apoptosis-related proteins were investigated using three methods: TdT-mediated dUTP digoxigenin nick end labeling (TUNEL) technique, active cysteine -dependant aspartate specific proteases (caspase)-3 and poly ADP-ribose polymerase (PARP) in coronary arteries of human subjects ranging from 25 to 92 years. We found no significant correlation between age and the apoptotic index using the three methods. The percentage of active caspase-3 positive cells was found to be significantly higher in men than in women (9.11 ± 12.3 cells/mm 2 versus 2.01 ± 4.55 cells/mm 2, respectively, p = 0.017). These sex-related differences did not reach statistical significance using TUNEL (9.93 ± 17 and 2.61 ± 4.58 cells/mm 2, p = 0.32) and PARP methods (3.42 ± 7.74 and 0.86 ± 0.95 cells/mm 2, p < 0.49). This is the first report of detection of apoptotic cells in the human arterial wall in adult subjects free from arterial diseases. Apoptosis is an attractive hypothesis to account for organ aging, but our study suggests that apoptosis is not a key factor in aging of the arterial wall.

Measurement of the ultrasonic properties of human coronary arteries in vitro with a 50-MHz acoustic microscope

Ultrasonic attenuation coefficient, wave propagation speed and integrated backscatter coefficient (IBC) of human coronary arteries were measured in vitro over the -6 dB frequency bandwidth (36 to 67 MHz) of a focused ultrasound transducer (50 MHz, focal distance 5.7 mm, f/number 1.7). Corrections were made for diffraction effects. Normal and diseased coronary artery sub-samples (N = 38) were obtained from 10 individuals at autopsy. The measured mean +/- SD of the wave speed (average over the entire vessel wall thickness) was 1581.04 +/- 53.88 m/s. At 50 MHz, the average attenuation coefficient was 4.99 +/- 1.33 dB/mm with a frequency dependence term of 1.55 +/- 0.18 determined over the 36- to 67-MHz frequency range. The IBC values were: 17.42 +/- 13.02 (sr.m)-1 for thickened intima, 11.35 +/- 6.54 (sr.m)-1 for fibrotic intima, 39.93 +/- 50.95 (sr.m)-1 for plaque, 4.26 +/- 2.34 (sr.m)-1 for foam cells, 5.12 +/- 5.85 (sr.m)-1 for media and 21.26 +/- 31.77 (sr.m)-1 for adventitia layers. The IBC results indicate the possibility for ultrasound characterization of human coronary artery wall tissue layer, including the situations of diseased arteries with the presence of thickened intima, fibrotic intima and plaque. The mean IBC normalized with respect to the mean IBC of the media layer seems promising for use as a parameter to differentiate a plaque or a thickened intima from a fibrotic intima.

Noninvasive in vivo human coronary artery lumen and wall imaging using black-blood magnetic resonance imaging.

High-resolution MRI has the potential to noninvasively image the human coronary artery wall and define the degree and nature of coronary artery disease. Coronary artery imaging by MR has been limited by artifacts related to blood flow and motion and by low spatial resolution. We used a noninvasive black-blood (BB) MRI (BB-MR) method, free of motion and blood-flow artifacts, for high-resolution (down to 0.46 mm in-plane resolution and 3-mm slice thickness) imaging of the coronary artery lumen and wall. In vivo BB-MR of both normal and atherosclerotic human coronary arteries was performed in 13 subjects: 8 normal subjects and 5 patients with coronary artery disease. The average coronary wall thickness for each cross-sectional image was 0.75+/-0.17 mm (range, 0.55 to 1.0 mm) in the normal subjects. MR images of coronary arteries in patients with >/=40% stenosis as assessed by x-ray angiography showed localized wall thickness of 4.38+/-0.71 mm (range, 3.30 to 5.73 mm). The difference in maximum wall thickness between the normal subjects and patients was statistically significant (P<0.0001). In vivo high-spatial-resolution BB-MR provides a unique new method to noninvasively image and assess the morphological features of human coronary arteries. This may allow the identification of atherosclerotic disease before it is symptomatic. Further studies are necessary to identify the different plaque components and to assess lesions in asymptomatic patients and their outcomes.

The expression of beta 1 integrins in human coronary artery.

The beta 1 integrin adhesion receptors mediate the binding of cells to extracellular matrices, facilitating their growth, migration, and capacity to deposit matrix proteins: important factors in arterial restenosis and atherosclerosis. The expression of integrins in human coronary artery is, however, unexplored. The aim of the current study was, therefore, to define the expression of beta 1 integrins by cultured human coronary artery vascular smooth muscle cells (hCAVSMC) and in normal human coronary artery; confirming whether or not this differs from the repertoire found in other species and human vessels. The expression of beta 1 integrins by hCAVSMC was assessed by immuno-precipitation and the alkaline phosphatase anti-alkaline phosphatase (APAAP) immunochemical technique. In addition, mRNA expression was defined by reverse transcription polymerase chain reaction (RT-PCR). Normal adult human coronary arteries (n = 4) were also stained by the APAAP method. In vitro hCAVSMC express alpha 2 beta 1 (a collagen and occasional laminin receptor) and alpha 5 beta 1 (a fibronectin receptor) with lesser expression of alpha 3 beta 1 (a multifunctional receptor). They do, however, possess mRNA for several other integrins. Cells within the media of human coronary artery wall express alpha 3 beta 1 and alpha 5 beta 1 but not alpha 2 beta 1: instead the alternative collagen/laminin receptor, alpha 1 beta 1, is expressed in vivo. This pattern of expression differs subtly from that described in rats through it closely parallels that found in other human arteries.

Coronary arterial dimensions and cell populations in ageing man

A combined computerised morphometric and immunocytochemical study on the ageing human coronary arterial wall shows that dilatation of the vessel is related to age but is less related to the degree of atherosclerosis. Loss of medial smooth muscle is not related to either atherosclerosis or dilatation of the artery. Occasional cases with severe medial and adventitial infiltrates of inflammatory cells (mainly T-cells and macrophages) do develop marked ectasia, but this arteritis is rather rare and, thus, an infrequent cause of dilatation.

Attenuation of the media of coronary arteries in advanced atherosclerosis

Human coronary artery wall architecture was analyzed in detail in 127 histologic sections with varying degrees of narrowing due to atherosclerotic plaque. A planimetry-microscope system was used to morphometrically determine percent luminal cross-sectional area narrowing due to atherosclerotic plaque, absolute area of the coronary artery media and total cross-sectional area of the coronary artery section. In 65 sections in which the native coronary artery lumen was narrowed less than 75%, the area of the coronary artery media corrected for total coronary cross-sectional area (Mc) was 0.244 +/- 0.055 mm2. In contrast, among 62 sections in which the coronary artery lumen was narrowed more than 75% in cross-sectional area, Mc measured 0.180 +/- 0.078 (p less than 0.001). Thus, in coronary artery segments with advanced atherosclerosis, there is substantial attenuation of the media, normally the principal component of the coronary artery wall.

泡沫細胞と粥腫退縮

The appearance of the human aortic wall and coronary arterial wall was examined by Scanning (SEM), Transmission electron microscopy (TEM) and chemical analysis in order to clarify the mechanism of removal of lipids deposited in the aorta.The findings of SEM and TEM showed that circulating liucocytes penetrated into the arterial wall through endotherial marginal folds in the early atherosclerotic lesion. In some specimens many spherical bodies with a diameter of 10-30μ were observed in the subendotherial space and intimal media. After the leucocytes had phagotized a large quantity of the lipids in the arterial wall, they were transformed into lipid-including cells, which we named “lipid-containers.” Furthermore, we found that some lipid-containers returned from the subendotherial space into the blood stream. Possibly, many lipids in the atherosclerotic lesion can be removed by this process. From those new findings, it is considered that the leucocytes (macrophage) play an important role in regression and prevention of atherosclerosis of human arteries at the cellular level. The human arterial wall was divided into three sections such as intima, intimal media and media by cryostat.The small lipids particles in the macrophages were isolated by ultracentrifugation after the three sections were digested with proteolytic enzymes. The lipid particles isolated from intima were small in diameter and very low density (1.006). But those from media were big and high density (1.210). The latter could not be so easily removed from arterial wall.From above findings, it suggested that mobility of macrophages played an important role of lipids deposition in the arterial wall.

Effect of age on the mechanical properties and biochemical composition of human cardiac arteries

1. The human left coronary arterial wall in subjects from age 15 to 28 has better deformative properties than the right artery.