Di Chang

Image-guided Pro-angiogenic Therapy in Diabetic Stroke Mouse Models Using a Multi-modal Nanoprobe

Purpose: The efficacy of pro-angiogenic therapy is difficult to evaluate with current diagnostic modalities. The objectives were to develop a non-invasive imaging strategy to define the temporal characteristics of angiogenesis and to evaluate the response to pro-angiogenic therapy in diabetic stroke mouse models. Methods: A home-made ανβ3 integrin-targeted multi-modal nanoprobe was intravenously injected into mouse models at set time points after photothrombotic stroke. Magnetic resonance imaging (MRI) and near-infrared fluorescence (NIRF) imaging were carried out at 24 h post-injection. Bone marrow-derived endothelial progenitor cells (EPCs) were infused into the mouse models of ischemic stroke to stimulate angiogenesis. Results: The peak signal intensity in the ischemic-angiogenic area of diabetic and wild-type mouse models was achieved on day 10, with significantly lower signal enhancement observed in the diabetic models. Although the signal intensity was significantly higher after EPC treatment in both models, the enhancement was less pronounced in the diabetic animals compared with the wild-type controls. Histological analysis revealed that the microvessel density and expression of β3 integrin were correlated with the signal intensity assessed with MRI and NIRF imaging. Conclusions: The non-invasive imaging method could be used for early and accurate evaluation of the response to pro-angiogenic therapy in diabetic stroke models.

Synergistic Effects of Transplanted Endothelial Progenitor Cells and RWJ 67657 in Diabetic Ischemic Stroke Models

Background and Purpose— An immature vascular phenotype in diabetes mellitus may cause more severe vascular damage and poorer functional outcomes after stroke, and it would be feasible to repair damaged functional vessels using endothelial progenitor cell (EPC) transplantation. However, high glucose induces p38 mitogen-activated protein kinase activation, which can accelerate the senescence and apoptosis of EPCs. The aim of this study was to investigate the combined effects of EPC transplantation and p38 mitogen-activated protein kinase inhibitor administration on diabetic stroke outcomes. Methods— Bone marrow–derived EPCs were injected intra-arterially into db/db mice after ischemic stroke induction. RWJ 67657 (RWJ), a p38 mitogen-activated protein kinase inhibitor, was administered orally for 7 consecutive days, with the first dose given 30 minutes before stroke induction. Functional outcome was determined at days 0, 1, 7, 14, and 21. Angiogenesis, neurogenesis, infarct volume, and Western blotting assays were performed on day 7, and white matter remodeling was determined on day 14. Results— Neither EPC transplantation nor RWJ administration alone significantly improved diabetic stroke outcome although RWJ displayed a potent anti-inflammatory effect. By both improving the functioning of EPCs and reducing inflammation, EPC transplantation plus RWJ administration in vivo synergistically promoted angiogenesis and neurogenesis after diabetic stroke. In addition, the white matter remodeling, behavioral scores, and expressions of vascular endothelial growth factor and brain-derived neurotrophic factor were significantly increased in diabetic mice treated with both EPCs and RWJ. Conclusions— The combination of EPC transplantation and RWJ administration accelerated recovery from diabetic stroke, which might have been caused by increased levels of proangiogenic and neurotrophic factors.

Salvaging brain ischemia by increasing neuroprotectant uptake via nanoagonist mediated blood brain barrier permeability enhancement.

Ischemic stroke is a leading cause of adult disability and cognitive impairment worldwide. Neuroprotective therapy aims to save neurons by impeding the deleterious ischemic insults. However, the low efficiency of the neuroprotectants crossing blood brain barrier (BBB) prevents their clinical translation. In this work, a nanoagonist (NA) was developed to enhance neuroprotectant uptake by specifically increasing BBB permeability in brain ischemia. This NA first targeted ischemic brain vasculatures, temporarily opened local BBB by activating adenosine 2A receptors, and up-regulated the neuroprotectant uptake in brain ischemia. This NA significantly increased the delivery of superoxide dismutase (SOD), a free radical scavenger, into mouse brain ischemia. The combined treatment of NA/SOD achieved a five-fold ischemic volume reduction rate compared to the animal models treated with SOD alone. Non-invasive magnetic resonance imaging (MRI) confirmed the ischemia targeted BBB opening, increased brain drug delivery efficiency and up-regulated therapeutic response during the combined NA/SOD treatment. Since the inefficient brain drug delivery is a general problem for the treatment of central nervous system (CNS) diseases, this work provides a novel strategy to deliver therapeutics by crossing BBB with high efficiency and targeting specificity.

Role of P38 MAPK on MMP Activity in Photothrombotic Stroke Mice as Measured using an Ultrafast MMP Activatable Probe.

Matrix metalloproteinases (MMPs) exert a dual effect in ischemic stroke and thus represent an ideal target for detection and therapy. However, to date, all clinical trials of MMP inhibitors have failed, and alternative drug candidates and therapeutic targets are urgently required. Nonetheless, further investigations are limited by the lack of non-invasive imaging techniques. Here, we report a novel, fast and ultrasensitive MMP activatable optical imaging probe for the dynamic visualization of MMP activity in photothrombotic stroke mice. This probe provides a significant signal enhancement in as little as 15 min, with the highest signal intensity occurring at 1 h post-injection, and shows high sensitivity in measuring MMP activity alterations, which makes it specifically suitable for the real-time visualization of MMP activity and drug discovery in preclinical research. Moreover, using this probe, we successfully demonstrate that the regulation of the p38 mitogen-activated protein kinase (MAPK) signal pathway is capable of modulating MMP activity after stroke, revealing a novel regulatory mechanism of postischemic brain damage and overcoming the limitations of traditional therapeutic strategies associated with MMP inhibitors by using a non-invasive molecular imaging method.

Significant perturbation in renal functional magnetic resonance imaging parameters and contrast retention for iodixanol compared with iopromide: an experimental study using blood-oxygen-level-dependent/diffusion-weighted magnetic resonance imaging and computed tomography in rats.

ObjectivesThe objective of this study was to investigate the renal changes after intravenous administration of a high dose of either iodixanol or iopromide using functional magnetic resonance imaging (MRI) and computed tomography (CT). Materials and MethodsThe study was approved by the institutional committee on animal research. Seventy-two male Sprague-Dawley rats were divided into 5 cohorts, comprising normal saline (NS), iopromide, iopromide + NS, iodixanol, and iodixanol + NS. Intravenous contrast was administrated at 8 g iodine/kg of body weight. Renal CT, quantitative functional MRI of blood-oxygen-level–dependent (BOLD) imaging and diffusion-weighted imaging (DWI), and histologic examinations were performed for 18 days after contrast administration. Statistical analysis was performed by using 1-way analysis of variance, Mann-Whitney test, and regression analysis. ResultsIn the renal cortex, BOLD showed persistent elevation of R2* and DWI showed persistent suppression of apparent diffusion coefficient after iodixanol administration for 18 days. Compared with iopromide, adjusted &Dgr;R2* (&Dgr;R2*adj) was significantly higher in the iodixanol group from 1 hour to 18 days (P < 0.04) after contrast; adjusted &Dgr;ADC (&Dgr;ADCadj) was significantly more pronounced at day 6 (P = 0.01) after contrast. The iodixanol cohort also exhibited persistently higher attenuation in the renal cortex on CT and more severe microscopic renal cortical vacuolization up to 18 days. Intravenous hydration decreased the magnetic resonance changes in both groups but more markedly with iodixanol. ConclusionsAt high doses, iodixanol induced greater changes in renal functional MRI (BOLD and DWI) relative to iopromide. Combined with longer contrast retention within the kidney, this suggests that iodixanol may produce more severe and longer-lasting contrast-induced renal damage.

Time-Efficient Myocardial Contrast Partition Coefficient Measurement from Early Enhancement with Magnetic Resonance Imaging

Objective Our purpose was to validate an early enhancement time point for accurately measuring the myocardial contrast partition coefficient (lambda) using dynamic-equilibrium magnetic resonance imaging. Materials and Methods The pre- and post-contrast longitudinal relaxation rates (reciprocal of T1) of the interventricular septum (R1m) and blood pool (R1b) were obtained from fifteen healthy volunteers and three diabetic patients with hypertension using two optimized T1 mapping sequences (modified Look-Locker inversion recovery) on a 3-Tesla magnetic resonance scanner. Reference lambda values were calculated as the slope of the regression line of R1m versus R1b at dynamic equilibrium (multi-point regression method). The simplified pre-/post-enhancement two-acquisition method (two-point method) was used to calculate lambda by relating the change in R1m and R1b using different protocols according to the acquisition stage of the post-enhancement data point. The agreement with the referential method was tested by calculating Pearsons correlation coefficient and the intra-class correlation coefficient. Results The lambda values measured by the two-point method increased (from 0.479±0.041 to 0.534±0.043) over time from 6 to 45 minutes after contrast and exhibited good correlation with the reference at each time point (r≥0.875, p<0.05). The intra-class correlation coefficient on absolute agreement with the reference lambda was 0.946, 0.929 and 0.922 at the 6th, 7th and 8th minutes and dropped from 0.878 to 0.403 from the 9th minute on. Conclusions The time-efficient two-point method at 6–8 minutes after the Gd-DTPA bolus injection exhibited good agreement with the multi-point regression method and can be applied for accurate lambda measurement in normal myocardium.

Noninvasive Identification of Renal Hypoxia in Experimental Myocardial Infarctions of Different Sizes by Using BOLD MR Imaging in a Mouse Model

Purpose To test the feasibility of using blood oxygen level-dependent (BOLD) magnetic resonance (MR) imaging to measure alterations in renal oxygenation in a mouse model with experimental myocardial infarctions (MIs) of different sizes. Materials and Methods The study was approved by the local animal ethics committee. One hundred eighty-nine male C57BL/6 J mice were randomly subjected to MI surgery (with different locations of left anterior descending coronary artery occlusion) or sham surgery, defined as the exposure of the heart but no ligation. Mice with MI underwent late gadolinium enhancement imaging 1 day after occlusion to confirm infarct size. Mice were sorted into three groups: those with large MI (n = 48), those with small MI (n = 48), and those with sham operation (n = 36). Renal BOLD MR imaging was performed before and 1, 7, 14, 28, and 60 days after MI, and histologic analysis of renal hypoxia-inducible factor-1α (HIF-1α) and kidney injury molecule-1 (KIM-1) was performed to evaluate tissue hypoxia and kidney injury in subgroups imaged at each time point. The relationships between the BOLD R2* and HIF-1α expression and between HIF-1α and KIM-1 expression were assessed. Statistical analyses were performed with one-way analysis of variance or the Kruskal-Wallis test and Spearman correlation test. Results A significant elevation in R2* was detected in the MI groups compared with the sham group in the cortex (P < .001 for large MI vs sham group; P = .007 for small MI vs sham group) and medulla (P < .001 for large MI vs sham group; P = .003 for small MI vs sham group) on day 60, and R2* was higher in the large MI group than in the small MI group (P < .001). Renal HIF-1α expression was increased after MI and showed linear correlation with R2* in the cortex (R2 = 0.56) and medulla (R2 = 0.63). In addition, an increase in renal KIM-1 was observed in the MI groups compared with the sham group on day 60 (sham group, 53.9 × 103 arbitrary units [au] ± 35.2; large MI group, 389.3 × 103 au ± 99.8; and small MI group, 185.8 × 103 au ± 91.9; P < .001 for large MI group vs sham group; P = .037 for small MI group vs sham group), and renal KIM-1 showed a positive correlation with HIF-1α (R2 = 0.68). Conclusion The magnitude of renal hypoxia with MIs of different sizes can be noninvasively measured with BOLD MR imaging, and increased renal hypoxia is a potential risk factor for progressive tubulointerstitial injury in mouse kidneys.

Correlations between the abdominal fat-related parameters and severity of coronary artery disease assessed by computed tomography

Background Obesity is considered a challenging public problem, which has been proven to be closely associated with coronary artery disease (CAD). Each risk factor of CAD has been separately studied many times in the past, but very few have comprehensively and quantitatively evaluated the relationship between the abdominal fat-related parameters and severity of CAD. The aim of this study was to analyze whether the abdominal fat-related parameters were associated with severity of CAD using abdominal non-enhanced computed tomography (NECT). Methods Patients who went through both abdominal NECT and invasive coronary angiography (ICA) were included and retrospectively analyzed. The abdominal fat-related parameters [the ratio of visceral adipose tissue to subcutaneous adipose tissue (VAT/SAT ratio)] and traditional cardiovascular risk factors were evaluated in the participants with or without obstructive CAD. The correlations between the abdominal fat-related parameters and severity of CAD were assessed, and the multivariable logistic regression analysis was performed to investigate the parameters that could be used to predict the severity of CAD. Results A total of 223 consecutive subjects (obstructive CAD group, n=117; non-obstructive CAD group, n=106) were analyzed. The VAT/SAT ratio was significantly higher (0.95±0.33 vs. 0.70±0.25, P<0.001) in obstructive CAD (O-CAD) patients than that in non-obstructive CAD (NO-CAD) patients. There was a trend to having nonalcoholic fatty liver disease (NAFLD) in the O-CAD patients than that of NO-CAD (P=0.002); the abdominal aortic calcification (AAC) score in O-CAD patients were higher than that in NO-CAD patients (P<0.001). The multivariable logistic regression analysis demonstrated that VAT/SAT ratio, NAFLD, and AAC score were independent predictors of O-CAD after adjusting the traditional cardiovascular risk factors. The area under the curve (AUC) of the combination of the above risk factors is 0.85, which leads to an increase in AUC than each risk factor alone in differentiating patients with or without O-CAD. Conclusions VAT/SAT ratio, NAFLD, and AAC score are correlated with the severity of CAD, indicating their characteristics of being independent risk factors for O-CAD, irrespective of the traditional cardiovascular risk factors. Those CT-derived parameters may make positive contributions to the differentiation of the patients with increased risk of O-CAD.