Home Atherosclerotic Lesions Rich in Macrophages or Smooth Muscle Cells Discriminated in Rabbit Iliac Arteries Based on T1 Relaxation Time and Lipid Content
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Atherosclerotic Lesions Rich in Macrophages or Smooth Muscle Cells Discriminated in Rabbit Iliac Arteries Based on T1 Relaxation Time and Lipid Content

Rationale and Objectives

Atherothrombosis usually occurs on macrophage- and lipid-rich unstable plaque, but rarely on smooth muscle cell (SMC)-rich stable plaque. Magnetic resonance imaging (MRI) has been extensively applied for noninvasive vascular imaging. We therefore investigated whether MRI provides valuable information about the characteristics of atherosclerotic vessels using rabbit models of macrophage-rich or SMC-rich atherosclerotic arteries.

Materials and Methods

Rabbits were fed with a conventional (CD group, n = 3) or 0.5% cholesterol (ChD group, n = 3) diet for 1 week before and 3 weeks after balloon injury of the left iliac arteries. Three weeks later, these arteries were investigates by 1.5 T MRI and by conventional angiographic imaging, followed by histological and immunohistochemical analyses.

Results

Three weeks after balloon injury, injured iliac arteries of both groups formed neointima with luminal stenosis. Conventional and MRI angiographic findings of the luminal diameter significantly and positively correlated. T1 relaxation time was significantly shorter and the lipid content was much higher in injured arteries from the ChD than from the CD group. The injured arteries from the ChD also contained more macrophages and less SMCs that those from the CD group. The T1 relaxation time and lipid content in injured arteries negatively and positively correlated with the degree of macrophage accumulation, respectively.

Conclusion

These results showed that MRI could provide valuable information about luminal stenosis and the characteristics of atherosclerotic vessels in rabbits.

Thrombus formation after atherosclerotic plaque disruption is a major cause of acute coronary syndrome and stroke. Atherothrombosis usually occurs on macrophage- and lipid-rich unstable plaque, but rarely on smooth muscle cell (SMC)-rich stable plaque . Beside luminal stenosis, plaque composition and morphology are key determinants of whether a plaque will cause cardiovascular events. Despite major advances in treatment, a high ratio of apparently healthy and asymptomatic patients dies of atherosclerosis . Therefore the development of screening and diagnostic methods to identify and characterize plaque composition is very important to determine the patient-specific risk of cardiovascular events and improve treatment strategies.

Magnetic resonance imaging (MRI) is used extensively for noninvasive vascular imaging, because it can identify components of atherosclerotic plaques such as a lipid-rich/necrotic core, calcification, and hemorrhage . It can also visualize and quantify lipid-rich human carotid plaque and the rabbit aorta . However, whether or not MRI can distinguish between stable and unstable atherosclerotic vessels in vivo is unknown. The aim of the study is to investigate whether MRI can distinguish between rabbit iliac arteries with macrophage-rich and SMC-rich atherosclerotic lesions.

Materials and methods

Experimental Design

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MRI

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Ex vivo Study

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Image and Data Analysis

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T1=TRln(sinα⋅cosβ⋅Iβ−sinβ⋅cosα⋅Iαsinα⋅Iβ−sinβ⋅Iα) T

1

=

T

R

ln

(

sin

α

cos

β

I

β

sin

β

cos

α

I

α

sin

α

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sin

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T2=TE2−TE1ln(I1I2) T

2

=

T

E

2

T

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1

ln

(

I

1

I

2

)

Lipid content by percentage (%) was derived from the following respective equations :

Lipid content(%)=I(T1WIwithoutfatsaturation)−I(T1WIwithfatsaturation)I(T1WIwithoutfatsaturation)×100 Lipid content

(

%

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=

I

(

T

1

W

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without

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(

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with

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100

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Serum Lipid Marker Sampling and Analysis

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Light Microscopy and Immunohistochemistry

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Statistical Analysis

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Results

Serum Concentrations of TC and TG 4 Weeks after Diets

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Magnetic Resonance Angiography 3 Weeks after Balloon Injury

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Figure 1, Representative magnetic resonance (MR) angiograms of rabbit iliac arteries. MR angiograms of the conventional cholesterol group (CD) (a) and 0.5% cholesterol group (ChD) (b) rabbits. Injured iliac arteries (left) of both groups have a narrower lumen (arrows) than uninjured arteries. (c) Luminal diameters of uninjured arteries (open box) and 3 weeks (dotted box) after balloon injury. ( n = 6 each; ∗ P < .001). Data are presented as medians (horizontal bar) , quartile ranges (boxes) , and 90th percentiles (error bars) . (d) Linear regression analysis of associations between conventional and MR angiography with respect to luminal diameter.

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MRI in vivo 3 Weeks after Balloon Injury

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Figure 2, In vivo axial magnetic resonance (MR) images and MR parameters of rabbit iliac arteries. (a) Luminal narrowing appears as thicker walls in injured arteries (arrows) of both groups compared with uninjured arteries. Thickened artery of 0.5% cholesterol (ChD) rabbit shows low- or high-signal intensity in T1-weighted image with or without fat saturation, respectively. (b) T1 and T2 relaxation times and lipid contents of iliac arteries in the conventional cholesterol (CD) and ChD groups, uninjured arteries (open box) and 3 weeks after balloon injury (dotted box) ( n = 6 each; ∗ P < .001). Data are shown as medians (horizontal bar) , quartile ranges (boxes) , and 90th percentiles (error bars) .

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Histopathological Correlation

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Figure 3, Representative microphotographs and linear regression analyses of iliac arteries 3 weeks after balloon injury. (a) Microphotographs of injured iliac arteries in both groups. Neointima has formed in injured iliac arteries. N: neointima; M: media; Ad: adventitia. (b) Immunopositive areas for RAM11 and HHF35 of iliac arteries 3 weeks after balloon injury in both groups ( n = 6 each; ∗ P < .05). Data are presented as medians (horizontal bar) , quartile ranges (boxes) , and 90th percentiles (error bars) . (c) Linear regression analysis of associations between relaxation times and immunopositive areas in injured iliac arteries.

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Ex vivo MRI and Histopathological Correlation

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Figure 4, Magnetic resonance parameters ex vivo and linear regression analyses. (a) T1 and T2 relaxation times, and lipid contents of iliac arteries in the conventional cholesterol group (CD) (open box) and 0.5% cholesterol (ChD) (dotted box) groups at 3 weeks after balloon injury ( n = 8 each; ∗ P < .001). (b) Immunopositive areas for RAM11 and HHF35 of iliac arteries at 3 weeks after balloon injury in both groups ( n = 8 each; ∗ P < .001). Data are presented as medians (horizontal bars) , quartile ranges (boxes) , and 90th percentiles (error bars) . (c) Linear regression analysis of associations between relaxation times and immunopositive area in injured iliac arteries.

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Discussion

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