Gezim Bala

Echocardiographic integrated backscatter for detecting progression and regression of aortic valve calcifications in rats

BackgroundCalcification is an independent predictor of mortality in calcific aortic valve disease (CAVD). The aim of this study was to evaluate the use of non-invasive, non-ionizing echocardiographic calibrated integrated backscatter (cIB) for monitoring progression and subsequent regression of aortic valvular calcifications in a rat model of reversible renal failure with CAVD, compared to histology.Methods28 male Wistar rats were prospectively followed during 21 weeks. Group 1 (N=14) was fed with a 0.5% adenine diet for 9 weeks to induce renal failure and CAVD. Group 2 (N=14) received a standard diet. At week 9, six animals of each group were killed. The remaining animals of group 1 (N=8) and group 2 (N=8) were kept on a standard diet for an additional 12 weeks. cIB of the aortic valve was calculated at baseline, 9 and 21 weeks, followed by measurement of the calcified area (Ca Area) on histology.ResultsAt week 9, cIB values and Ca Area of the aortic valve were significantly increased in the adenine-fed rats compared to baseline and controls. After 12 weeks of adenine diet cessation, cIB values and Ca Area of group 1 decreased compared to week 9, while there was no longer a significant difference compared to age-matched controls of group 2.ConclusionscIB is a non-invasive tool allowing quantitative monitoring of CAVD progression and regression in a rat model of reversible renal failure, as validated by comparison with histology. This technique might become useful for assessing CAVD during targeted therapy.

Animal models of organic heart valve disease

Heart valve disease is a frequently encountered pathology, related to high morbidity and mortality rates in industrialized and developing countries. Animal models are interesting to investigate the causality, but also underlying mechanisms and potential treatments of human valvular diseases. Recently, animal models of heart valve disease have been developed, which allow to investigate the pathophysiology, and to follow the progression and the potential regression of disease with therapeutics over time. The present review provides an overview of animal models of primary, organic heart valve disease: myxoid age-related, infectious, drug-induced, degenerative calcified, and mechanically induced valvular heart disease.

Occurrence of cardiovascular calcifications in normal, aging rats.

BACKGROUND Cardiovascular calcification is an independent predictor of morbidity and mortality and increases with age. Animal models are frequently used to investigate the underlying pathophysiology. Only scarce data regarding the effect of aging on calcifications in these animal models are available. The aim of this study is to investigate the occurrence of cardiovascular calcifications in normal, aging rats. METHODS A mixed inbred/outbred population of 44 male Lewis/Wistar rats was studied. Group 1 of three-month-old rats, group 2 twelve-month-old, group 3 twenty-four-month-old and group 4 thirty-month-old rats. Calibrated integrated backscatter (cIB) values and blood parameters (creatinine, parathyroid hormone (PTH)) were measured, followed by ex-vivo micro-CT and histology as reference methods. RESULTS Cardiovascular calcifications developed with age, as demonstrated by significantly increasing cIB values of the aortic valve and myocardium. This was confirmed by a significant increase in the calcified volume on ex-vivo micro-CT and in the histological calcium score. There was also a significantly higher level of creatinine and PTH with age. CONCLUSIONS As in humans, cardiovascular calcifications progressively increase with age in the normal rat. Therefore the aging rat model could be used for studying calcifying cardiovascular disease. cIB might have a value in future studies for the early detection of subclinical calcifications in humans.

Recent Advances in Visualizing Vulnerable Plaque: Focus on Noninvasive Molecular Imaging

Traditional imaging methods in atherosclerosis have focused primarily on anatomic information. Imaging approaches that visualize molecular targets rather than anatomic structures may emphasize biologic aspects of atherosclerosis. Molecular imaging of atherosclerotic lesions has become a crucial experimental tool and is now emerging in the clinical arena. In this review, we briefly highlight the rationale and fundamental principles of molecular imaging. We then discuss the promising imaging modalities, along with their potential limitations, and the molecular targets being investigated in experimental research. Finally, we summarize the most important clinical studies recently performed in humans.

Quantification of Calcium Amount in a New Experimental Model: A Comparison between Ultrasound and Computed Tomography

Purpose Calcification is an important prognostic factor in aortic valve stenosis. However, there is no ultrasound (US) method available to accurately quantify calcification in this setting to date. We aimed to validate a new US method for measuring the amount of calcium in an in vitro model, and compare it to computed tomography (CT), the current imaging gold standard. Materials and Methods An agar phantom (2% agar) was made, containing 9 different amounts of calcium-hydroxyapatite Ca5(PO4)3OH (2 to 50mg). The phantoms were imaged with micro-CT and US (10 MHz probe). The calcium area (areacalcium) and its maximum pixel value (PVmax) were obtained. These values were summed to calculate CT and US calcium scores (∑(areacalcium × PVmax)) and volumes (∑areacalcium). Both US- and CT-calcium scores were compared with the calcium amounts, and with each other. Results Both calcium scores correlated significantly with the calcium amount (R2 = 0.9788, p<0.0001 and R2 = 0.8154, p<0.0001 for CT and US respectively). Furthermore, there was a significant correlation between US and CT for calcium volumes (R2 = 0.7392, p<0.0001) and scores (R2 = 0.7391, p<0.0001). Conclusion We developed a new US method that accurately quantifies the amount of calcium in an in vitro model. Moreover it is strongly correlated with CT.

Translating Molecular Imaging of the Vulnerable Plaque—a Vulnerable Project?

Translating Molecular Imaging of the Vulnerable Plaque—a Vulnerable Project? Gezim Bala, Alexis Broisat, Tony Lahoutte, Sophie Hernot 1 In vivo Cellular and Molecular Imaging (ICMI/BEFY), Vrije Universiteit Brussel, Brussels, Belgium Department of Cardiology, UZ Brussel, Brussels, Belgium Radiopharmaceutiques Biocliniques, INSERM, 1039-Université de Grenoble, La Tronche, France Department of Nuclear Medicine, UZ Brussel, Brussels, Belgium

Echocardiographic integrated backscatter for the differentiation between aortic valve calcification and valvular myxoid degeneration in rats

AIMS Calcification is an independent predictor of mortality in aortic valve (AV) stenosis. Echocardiographic calibrated integrated backscatter (cIB) is a promising parameter for quantifying AV calcification. However, the ability of cIB to differentiate between calcification and valvular thickening has been questioned. Therefore, we aimed to use cIB to study AV calcification compared with non-calcified AV thickening in rats, with histology as reference. METHODS AND RESULTS Twenty male Wistar rats were studied. Group 1 (N = 6) received subcutaneous (SC) serotonin injections (60 mg/kg/day) for 12 weeks to induce myxoid non-calcified AV thickening. Group 2 (N = 7) received vitamin D3 (25,000 UI/kg/day) SC to induce AV calcification, and Group 3 (N = 7) received only vehicle SC for 10 weeks. cIB of the AV was calculated at the end of the study, followed by measurement of the percentage of the histological AV calcification. At the end of the study, cIB values and calcification percentages were significantly higher in vitamin D3-injected rats compared with serotonin-injected rats and controls. There was no significant difference in cIB values between serotonin-injected rats and controls (vitamin D3: 21.5 ± 3.0 dB*; serotonin: 11.8 ± 3.1 dB; control: 10.3 ± 3.4 dB; *P < 0.05). The percentage of histological calcification was significantly higher in the vitamin D3 group compared with the other groups. Serotonin-injected rats developed significant AV thickening. CONCLUSION Increased cIB values of the AV are related to increased calcification at histology and not to myxoid non-calcified valvular thickening. Therefore, cIB may be considered as a sensitive technique to quantify calcification of AV rather than for detecting non-calcified valvular thickening.

Radiometal-labeled anti-VCAM-1 nanobodies as molecular tracers for atherosclerosis – impact of radiochemistry on pharmacokinetics

Abstract Radiolabeling of nanobodies with radiometals by chelation has the advantage of being simple, fast and easy to implement in clinical routine. In this study, we validated 68Ga/111In-labeled anti-VCAM-1 nanobodies as potential radiometal-based tracers for molecular imaging of atherosclerosis. Both showed specific targeting of atherosclerotic lesions in ApoE−/− mice. Nevertheless, uptake in lesions and constitutively VCAM-1 expressing organs was lower than previously reported for the 99mTc-labeled analog. We further investigated the impact of different radiolabeling strategies on the in vivo biodistribution of nanobody-based tracers. Comparison of the pharmacokinetics between 68Ga-, 18F-, 111In- and 99mTc-labeled anti-VCAM-1 nanobodies showed highest specific uptake for 99mTc-nanobody at all time-points, followed by the 68Ga-, 111In- and 18F-labeled tracer. No correlation was found with the estimated number of radioisotopes per nanobody, and mimicking specific activity of other radiolabeling methods did not result in an analogous biodistribution. We also demonstrated specificity of the tracer using mice with a VCAM-1 knocked-down phenotype, while showing for the first time the in vivo visualization of a protein knock-down using intrabodies. Conclusively, the chosen radiochemistry does have an important impact on the biodistribution of nanobodies, in particular on the specific targeting, but differences are not purely due to the tracer’s specific activity.

Interaction of renal failure and dyslipidaemia in the development of calcific aortic valve disease in rats

Abstract Objective: Calcific aortic valve disease (CAVD) is currently the most common heart valve disease worldwide and is known to be an active process. Both renal failure and dyslipidaemia are considered to be promoting factors for the development of valvular calcifications. The aim of this study is to prospectively evaluate the respective contribution and interaction of renal failure and dyslipidaemia on CAVD in a rat model, using echocardiography and compared with histology. Methods and results: Sixty-eight male Wistar rats were prospectively divided in eight groups, each fed a different diet to induce renal failure alone and combined with hyperlipidaemia or hypercholesterolemia. CAVD was detected and quantified by calibrated integrated backscatter of ultrasound (cIB) and compared with the histological calcium score. The study follow-up was 20 weeks. At the end of the study, the cIB value and the calcium score of the aortic valve were significantly increased in the group with isolated renal failure but not with dyslipidaemia. The combination of renal failure with high cholesterol or high-fat diet did not significantly increase calcifications further. Conclusions: Renal failure alone does induce aortic valve calcifications in a rat model of CAVD, whereas dyslipidaemia alone does not. The combination of renal failure with dyslipidaemia does not increase calcification further. These findings suggest that a combination of atherosclerotic and calcifying factors is not required to induce aortic valve calcifications in this model.