Tuan Pham

Effect of PEG molecular weight on stability, T2 contrast, cytotoxicity, and cellular uptake of superparamagnetic iron oxide nanoparticles (SPIONs)

Superparamagnetic iron oxide nanoparticles (SPIONs) are currently unavailable as MRI contrast agents for detecting atherosclerosis in the clinical setting because of either low signal enhancement or safety concerns. Therefore, a new generation of SPIONs with increased circulation time, enhanced image contrast, and less cytotoxicity is essential. In this study, monodisperse SPIONs were synthesized and coated with polyethylene glycol (PEG) of varying molecular weights. The resulting PEGylated SPIONs were characterized, and their interactions with vascular smooth muscle cells (VSMCs) were examined. SPIONs were tested at different concentrations (100 and 500 ppm Fe) for stability, T2 contrast, cytotoxicity, and cellular uptake to determine an optimal formulation for in vivo use. We found that at 100 ppm Fe, the PEG 2K SPIONs showed adequate stability and magnetic contrast, and exhibited the least cytotoxicity and nonspecific cellular uptake. An increase in cell viability was observed when the SPION-treated cells were washed with PBS after 1h incubation compared to 5 and 24h incubation without washing. Our investigation provides insight into the potential safe application of SPIONs in the clinic.

Regions of Low Endothelial Shear Stress Colocalize With Positive Vascular Remodeling and Atherosclerotic Plaque Disruption An In Vivo Magnetic Resonance Imaging Study

Background— Local hemodynamic factors, particularly low endothelial shear stress (ESS), play a role in the focal formation of atherosclerosis. We used in vivo MRI to investigate the role of the magnitude of ESS on vascular remodeling, plaque burden, and disruption using a rabbit model of controlled atherothrombosis. Methods and Results— Atherosclerosis was induced in New Zealand white rabbits by cholesterol diet and endothelial denudation. MRI was performed before (pretrigger) and after (posttrigger) inducing plaque disruption with Russell viper venom and histamine. Of the 134 vascular segments studied, 28 contained thrombus (disrupted) and 106 did not (nondisrupted). Disrupted plaques were histologically characterized by a thin, inflamed fibrous cap, a dense lipid core, and mural thrombus. Pretriggered MRI revealed that disrupted plaques clustered at regions with low mean ESS (11.55±5.3 versus 20.9±9.74 dynes/cm2; P<0.001) and low peak ESS (21.5±11.2 versus 49.2±21.5 dynes/cm2; P<0.001) compared with nondisrupted plaques. The peak ESS negatively correlated with the plaque area (r=−0.56, P<0.001) and remodeling ratio (r=−0.4, P=0.008). There was also a negative correlation between the mean ESS and the remodeling ratio (r=−0.55, P<0.001). Both the peak ESS and the mean ESS did not correlate with the % stenosis; there was a weak but statistically significant correlation with the % cross-sectional narrowing (r=0.3, P=0.002 and r=0.2, P=0.04, respectively). Receiver operating characteristic analysis showed that both mean (Area under the curve=0.78; 95% CI, 0.69–0.87) and peak ESS (Area under the curve=0.85; 95% CI, 0.78–0.93) identified disrupted plaques. Conclusions— We demonstrated that low ESS is associated with plaque burden, positive vascular remodeling, and plaque disruption in a rabbit model. Assessment of ESS by noninvasive MRI might be useful for assessing atherosclerotic risk.

Identification of High-Risk Plaques by MRI and Fluorescence Imaging in a Rabbit Model of Atherothrombosis

Introduction The detection of atherosclerotic plaques at risk for disruption will be greatly enhanced by molecular probes that target vessel wall biomarkers. Here, we test if fluorescently-labeled Activatable Cell Penetrating Peptides (ACPPs) could differentiate stable plaques from vulnerable plaques that disrupt, forming a luminal thrombus. Additionally, we test the efficacy of a combined ACPP and MRI technique for identifying plaques at high risk of rupture. Methods and Results In an atherothrombotic rabbit model, disrupted plaques were identified with in vivo MRI and co-registered in the same rabbit aorta with the in vivo uptake of ACPPs, cleaved by matrix metalloproteinases (MMPs) or thrombin. ACPP uptake, mapped ex vivo in whole aortas, was higher in disrupted compared to non-disrupted plaques. Specifically, disrupted plaques demonstrated a 4.5~5.0 fold increase in fluorescence enhancement, while non-disrupted plaques showed only a 2.2~2.5 fold signal increase. Receiver operating characteristic (ROC) analysis indicates that both ACPPs (MMP and thrombin) show high specificity (84.2% and 83.2%) and sensitivity (80.0% and 85.7%) in detecting disrupted plaques. The detection power of ACPPs was improved when combined with the MRI derived measure, outward remodeling ratio. Conclusions Our targeted fluorescence ACPP probes distinguished disrupted plaques from stable plaques with high sensitivity and specificity. The combination of anatomic, MRI-derived predictors for disruption and ACPP uptake can further improve the power for identification of high-risk plaques and suggests future development of ACPPs with molecular MRI as a readout.

The influence of pericardial fat upon left ventricular function in obese females: evidence of a site-specific effect

BackgroundAlthough increased volume of pericardial fat has been associated with decreased cardiac function, it is unclear whether this association is mediated by systemic overall obesity or direct regional fat interactions. We hypothesized that if local effects dominate, left ventricular (LV) function would be most strongly associated with pericardial fat that surrounds the left rather than the right ventricle (RV).MethodsFemale obese subjects (n = 60) had cardiovascular magnetic resonance (CMR) scans to obtain measures of LV function and pericardial fat volumes. LV function was obtained using the cine steady state free precession imaging in short axis orientation. The amount of pericardial fat was determined volumetrically by the cardiac gated T1 black blood imaging and normalized to body surface area.ResultsIn this study cohort, LV fat correlated with several LV hemodynamic measurements including cardiac output (r = -0.41, p = 0.001) and stroke volume (r = -0.26, p = 0.05), as well as diastolic functional parameters including peak-early-filling rate (r = -0.38, p = 0.01), early late filling ratio (r = -0.34, p = 0.03), and time to peak-early-filling (r = 0.34, p = 0.03). These correlations remained significant even after adjusting for the body mass index and the blood pressure. However, similar correlations became weakened or even disappeared between RV fat and LV function. LV function was not correlated with systemic plasma factors, such as C-reactive protein (CRP), B-type natriuretic peptide (BNP), Interleukin-6 (IL-6), resistin and adiponectin (all p > 0.05).ConclusionsLV hemodynamic and diastolic function was associated more with LV fat as compared to RV or total pericardial fat, but not with systemic inflammatory markers or adipokines. The correlations between LV function and pericardial fat remained significant even after adjusting for systemic factors. These findings suggest a site-specific influence of pericardial fat on LV function, which could imply local secretion of molecules into the underlying tissue or an anatomic effect, both mechanisms meriting future evaluation.

Spatio-temporal texture (SpTeT) for distinguishing vulnerable from stable atherosclerotic plaque on dynamic contrast enhancement (DCE) MRI in a rabbit model

PURPOSE To develop a new spatio-temporal texture (SpTeT) based method for distinguishing vulnerable versus stable atherosclerotic plaques on DCE-MRI using a rabbit model of atherothrombosis. METHODS Aortic atherosclerosis was induced in 20 New Zealand White rabbits by cholesterol diet and endothelial denudation. MRI was performed before (pretrigger) and after (posttrigger) inducing plaque disruption with Russells-viper-venom and histamine. Of the 30 vascular targets (segments) under histology analysis, 16 contained thrombus (vulnerable) and 14 did not (stable). A total of 352 voxel-wise computerized SpTeT features, including 192 Gabor, 36 Kirsch, 12 Sobel, 52 Haralick, and 60 first-order textural features, were extracted on DCE-MRI to capture subtle texture changes in the plaques over the course of contrast uptake. Different combinations of SpTeT feature sets, in which the features were ranked by a minimum-redundancy-maximum-relevance feature selection technique, were evaluated via a random forest classifier. A 500 iterative 2-fold cross validation was performed for discriminating the vulnerable atherosclerotic plaque and stable atherosclerotic plaque on per voxel basis. Four quantitative metrics were utilized to measure the classification results in separating between vulnerable and stable plaques. RESULTS The quantitative results show that the combination of five classes of SpTeT features can distinguish between vulnerable (disrupted plaques with an overlying thrombus) and stable plaques with the best AUC values of 0.9631 ± 0.0088, accuracy of 89.98% ± 0.57%, sensitivity of 83.71% ± 1.71%, and specificity of 94.55% ± 0.48%. CONCLUSIONS Vulnerable and stable plaque can be distinguished by SpTeT based features. The SpTeT features, following validation on larger datasets, could be established as effective and reliable imaging biomarkers for noninvasively assessing atherosclerotic risk.

Regions of Low Endothelial Shear Stress Co-localize with Positive Vascular Remodeling and Atherosclerotic Plaque Disruption: An in vivo MRI Study

Background— Local hemodynamic factors, particularly low endothelial shear stress (ESS), play a role in the focal formation of atherosclerosis. We used in vivo MRI to investigate the role of the magnitude of ESS on vascular remodeling, plaque burden, and disruption using a rabbit model of controlled atherothrombosis. Methods and Results— Atherosclerosis was induced in New Zealand white rabbits by cholesterol diet and endothelial denudation. MRI was performed before (pretrigger) and after (posttrigger) inducing plaque disruption with Russell viper venom and histamine. Of the 134 vascular segments studied, 28 contained thrombus (disrupted) and 106 did not (nondisrupted). Disrupted plaques were histologically characterized by a thin, inflamed fibrous cap, a dense lipid core, and mural thrombus. Pretriggered MRI revealed that disrupted plaques clustered at regions with low mean ESS (11.55±5.3 versus 20.9±9.74 dynes/cm2; P<0.001) and low peak ESS (21.5±11.2 versus 49.2±21.5 dynes/cm2; P<0.001) compared with nondisrupted plaques. The peak ESS negatively correlated with the plaque area (r=−0.56, P<0.001) and remodeling ratio (r=−0.4, P=0.008). There was also a negative correlation between the mean ESS and the remodeling ratio (r=−0.55, P<0.001). Both the peak ESS and the mean ESS did not correlate with the % stenosis; there was a weak but statistically significant correlation with the % cross-sectional narrowing (r=0.3, P=0.002 and r=0.2, P=0.04, respectively). Receiver operating characteristic analysis showed that both mean (Area under the curve=0.78; 95% CI, 0.69–0.87) and peak ESS (Area under the curve=0.85; 95% CI, 0.78–0.93) identified disrupted plaques. Conclusions— We demonstrated that low ESS is associated with plaque burden, positive vascular remodeling, and plaque disruption in a rabbit model. Assessment of ESS by noninvasive MRI might be useful for assessing atherosclerotic risk.

Microbubbles as Theranostics Agents

Clinically, ultrasound (US) has been used as a cheap, quick, and effective form of imaging that provides information useful for diagnostic purposes. With the advent of microbubbles as US contrast agents, this simple imaging technique has evolved into a tool capable of providing molecularly targeted visualization of disease and controlled delivery of therapeutics. The simple, yet robust, structure of the microbubble allows for both internal and external modifications, which lead to a wide variety of clinical uses. This chapter will introduce the reader to microbubble fabrication, stabilization, drug loading, and targeting. The reader will also be briefly exposed to specific examples of current work done using microbubbles in areas of cancer treatment and protein/gene therapies. The work reviewed here is only a small fraction of the literature available on the subject matter and serves as an introduction to microbubbles as contrast agents, drug delivery vehicles, and theranostic particles.

Investigating the effect of fabrication method on the stability and acoustic response of microbubble agents

Microbubbles stabilized by a surfactant or polymer coating are already in clinical use as ultrasound imaging contrast agents. They have also been widely investigated as vehicles for drug delivery and gene therapy that can be tracked and triggered using ultrasound. Extensive studies have been made of the effects of the coating material and gas core on microbubble characteristics, but the influence of the fabrication method has received less attention. The aim of this study was to compare the behavior of microbubbles prepared using different techniques. Phospholipid-coated microbubbles were produced using sonication, electrospraying, or in a specially designed microfluidic device. The microbubbles were observed using optical, electron, and fluorescence lifetime imaging microscopy (FLIM) to interrogate their surface microstructure and stability over time. Their acoustic response was then determined in a flow chamber by detecting the pressure scattered from individual microbubbles as they passed through the f...

Regions of Low Endothelial Shear Stress Colocalize With Positive Vascular Remodeling and Atherosclerotic Plaque Disruption

Background— Local hemodynamic factors, particularly low endothelial shear stress (ESS), play a role in the focal formation of atherosclerosis. We used in vivo MRI to investigate the role of the magnitude of ESS on vascular remodeling, plaque burden, and disruption using a rabbit model of controlled atherothrombosis. Methods and Results— Atherosclerosis was induced in New Zealand white rabbits by cholesterol diet and endothelial denudation. MRI was performed before (pretrigger) and after (posttrigger) inducing plaque disruption with Russell viper venom and histamine. Of the 134 vascular segments studied, 28 contained thrombus (disrupted) and 106 did not (nondisrupted). Disrupted plaques were histologically characterized by a thin, inflamed fibrous cap, a dense lipid core, and mural thrombus. Pretriggered MRI revealed that disrupted plaques clustered at regions with low mean ESS (11.55±5.3 versus 20.9±9.74 dynes/cm2; P<0.001) and low peak ESS (21.5±11.2 versus 49.2±21.5 dynes/cm2; P<0.001) compared with nondisrupted plaques. The peak ESS negatively correlated with the plaque area (r=−0.56, P<0.001) and remodeling ratio (r=−0.4, P=0.008). There was also a negative correlation between the mean ESS and the remodeling ratio (r=−0.55, P<0.001). Both the peak ESS and the mean ESS did not correlate with the % stenosis; there was a weak but statistically significant correlation with the % cross-sectional narrowing (r=0.3, P=0.002 and r=0.2, P=0.04, respectively). Receiver operating characteristic analysis showed that both mean (Area under the curve=0.78; 95% CI, 0.69–0.87) and peak ESS (Area under the curve=0.85; 95% CI, 0.78–0.93) identified disrupted plaques. Conclusions— We demonstrated that low ESS is associated with plaque burden, positive vascular remodeling, and plaque disruption in a rabbit model. Assessment of ESS by noninvasive MRI might be useful for assessing atherosclerotic risk.

Regions of Low Endothelial Shear Stress Colocalize With Positive Vascular Remodeling and Atherosclerotic Plaque DisruptionClinical Perspective: An In Vivo Magnetic Resonance Imaging Study

Background— Local hemodynamic factors, particularly low endothelial shear stress (ESS), play a role in the focal formation of atherosclerosis. We used in vivo MRI to investigate the role of the magnitude of ESS on vascular remodeling, plaque burden, and disruption using a rabbit model of controlled atherothrombosis. Methods and Results— Atherosclerosis was induced in New Zealand white rabbits by cholesterol diet and endothelial denudation. MRI was performed before (pretrigger) and after (posttrigger) inducing plaque disruption with Russell viper venom and histamine. Of the 134 vascular segments studied, 28 contained thrombus (disrupted) and 106 did not (nondisrupted). Disrupted plaques were histologically characterized by a thin, inflamed fibrous cap, a dense lipid core, and mural thrombus. Pretriggered MRI revealed that disrupted plaques clustered at regions with low mean ESS (11.55±5.3 versus 20.9±9.74 dynes/cm2; P<0.001) and low peak ESS (21.5±11.2 versus 49.2±21.5 dynes/cm2; P<0.001) compared with nondisrupted plaques. The peak ESS negatively correlated with the plaque area (r=−0.56, P<0.001) and remodeling ratio (r=−0.4, P=0.008). There was also a negative correlation between the mean ESS and the remodeling ratio (r=−0.55, P<0.001). Both the peak ESS and the mean ESS did not correlate with the % stenosis; there was a weak but statistically significant correlation with the % cross-sectional narrowing (r=0.3, P=0.002 and r=0.2, P=0.04, respectively). Receiver operating characteristic analysis showed that both mean (Area under the curve=0.78; 95% CI, 0.69–0.87) and peak ESS (Area under the curve=0.85; 95% CI, 0.78–0.93) identified disrupted plaques. Conclusions— We demonstrated that low ESS is associated with plaque burden, positive vascular remodeling, and plaque disruption in a rabbit model. Assessment of ESS by noninvasive MRI might be useful for assessing atherosclerotic risk.