Comparison of small animal CT contrast agents

Non‐invasive in vivo small animal computed tomography (CT) imaging provides high resolution bone scans but cannot differentiate between soft tissues. For most applications injections of contrast agents (CAs) are necessary. Aim of this study was to uncover the advantages and disadvantages of commercially available CT CAs (ExiTron nano 12 000 and 6000, eXIA 160 and 160XL, Fenestra VC and LC) regarding their pharmacokinetics, toxicological side‐effects and the influence of anesthesia on the biodistribution, based on an injection volume of 100 μL/25 g body weight. The pharmacokinetics of the CAs were determined for up to five days. The CA‐induced toxicological/physiological side‐effects were evaluated by determining blood counts, liver enzymes, thyroxine and total protein values, pro‐inflammatory mediators (messenger ribonucleic acid (mRNA)), histology and immunohistochemistry. ExiTron nano 12 000 and 6000 yielded a long‐term contrast enhancement (CE) in the liver and spleen for up to five days. Some of the evaluated CAs did not show any CE at all. Anesthesia did not impair the CAs' biodistribution. The CAs differentially affected the body weight, blood counts, liver enzymes, thyroxine and total protein values. ExiTron nano 12 000 and 6000 induced histiocytes in the liver and spleen. Moreover, ExiTron nano 12 000 and eXIA 160 enhanced tumor necrosis factor (TNF) mRNA expression levels in the kidneys. Thus, we recommend ExiTron nano 12 000 and 6000 when multiple injections should be avoided. We recommend careful selection of the employed CA in order to achieve an acceptable CE in the organs of interest and to avoid influences on the animal physiology. Copyright © 2016 John Wiley & Sons, Ltd.     

Dendritic PARACEST contrast agents for magnetic resonance imaging

MRI contrast agents based on chemical exchange‐dependent saturation transfer (CEST), such as Yb(III)DOTAM complexes, are highly suitable for pH mapping. In this paper, the synthesis of Yb(III)DOTAM‐functionalized poly(propylene imine) dendrimers is described. The applicability of these dendritic PARACEST MRI agents for pH mapping has been evaluated on a 7 T NMR spectrometer and on a 3 T clinical MRI scanner. As expected, based on the different numbers of exchangeable amide protons, the lowest detectable concentration of the first and third generation dendritic PARACEST agents is by a respective factor of about 4 and 16 lower than that of a mononuclear reference complex. The pH dependence of the CEST effect observed for these compounds depends on the generation of the poly(propylene imine) dendrimer. Upon going to higher generations of the Yb(III)DOTAM‐terminated dendrimer, a shift of the maximum CEST effect towards lower pH values was observed. This allows for a fine‐tuning of the responsive pH region by varying the dendritic framework. Copyright © 2007 John Wiley & Sons, Ltd.     

Classification and basic properties of contrast agents for magnetic resonance imaging

A comprehensive classification of contrast agents currently used or under development for magnetic resonance imaging (MRI) is presented. Agents based on small chelates, macromolecular systems, iron oxides and other nanosystems, as well as responsive, chemical exchange saturation transfer (CEST) and hyperpolarization agents are covered in order to discuss the various possibilities of using MRI as a molecular imaging technique. The classification includes composition, magnetic properties, biodistribution and imaging applications. Chemical compositions of various classes of MRI contrast agents are tabulated, and their magnetic status including diamagnetic, paramagnetic and superparamagnetic are outlined. Classification according to biodistribution covers all types of MRI contrast agents including, among others, extracellular, blood pool, polymeric, particulate, responsive, oral, and organ specific (hepatobiliary, RES, lymph nodes, bone marrow and brain). Various targeting strategies of molecular, macromolecular and particulate carriers are also illustrated. Copyright © 2009 John Wiley & Sons, Ltd.     

Contrast enhancement by lipid‐based MRI contrast agents in mouse atherosclerotic plaques; a longitudinal study

The use of contrast‐enhanced MRI to enable in vivo specific characterization of atherosclerotic plaques is increasing. In this study the intrinsic ability of two differently sized gadolinium‐based contrast agents to enhance atherosclerotic plaques in ApoE^?/? mice was evaluated with MRI. We obtained a kinetic profile for contrast enhancement, as the literature data on optimal imaging time points is scarce, and assessed the longer‐term kinetics. Signal enhancement in the wall of the aortic arch, following intravenous injection of paramagnetic micelles and liposomes, was followed for 1 week. In vivo T₁‐weighted MRI plaque enhancement characteristics were complemented by fluorescence microscopy of NIR₆₆₄ incorporated in the contrast agents and quantification of tissue and blood Gd–DTPA. Both micelles and liposomes enhanced contrast in T₁‐weighted MR images of plaques in the aortic arch. The average contrast‐to‐noise ratio increased after liposome or micelle injection to 260 or 280% respectively, at 24 h after injection, compared with a pre‐scan. A second wave of maximum contrast enhancement was observed around 60–72 h after injection, which only slowly decreased towards the 1 week end‐point. Confocal fluorescence microscopy and whole body fluorescence imaging confirmed MRI‐findings of accumulation of micelles and liposomes. Plaque permeation of contrast agents was not strongly dependent on the contrast agent size in this mouse model. Our results show that intraplaque accumulation over time of both contrast agents leads to good plaque visualization for a long period. This inherent intraplaque accumulation might make it difficult to discriminate passive from targeted accumulation. This implies that, in the development of targeted contrast agents on a lipid‐based backbone, extensive timing studies are required. Copyright © 2012 John Wiley & Sons, Ltd.     

Magnetic resonance imaging of atherosclerosis by targeting extracellular matrix deposition with Gadofluorine M

As previously reported, Gadofluorine M‐enhanced magnetic resonance imaging clearly demarcates atherosclerotic plaques from the normal vessel wall. To date, the underlying mechanism has remained unknown. Gadofluorine M is a gadolinium‐containing macrocyclic contrast agent containing hydrophilic and hydrophobic moieties. To elucidate the mechanism of accumulation, fluorescently labeled and radioactively labeled derivates of Gadofluorine M were used to determine affinity and specificity of Gadofluorine M binding to blood serum and plaque components in vitro and for the distribution within the plaque of WHHL rabbits in vivo. Gadofluorine M binds to serum albumin, leading to a breakdown of micelles after intravenous injection. The affinity of Gadofluorine M to serum albumin is k_D = 2 µmol/l. Gadofluorine then penetrates the atherosclerotic plaque while bound to albumin and then accumulates within the extracellular, fibrous parts of the plaque by binding to collagens, proteoglycans and tenascin, having the same affinity to these plaque constituents as to albumin. In contrast, weak binding was determined to LDL (k_D = 2 mmol/l) and even no binding to hyaluronic acid. The driving force of binding and accumulation is the hydrophobic moiety of the molecules interacting with hydrophobic plaque materials. Thus, Gadofluorine M accumulates within the fibrous plaque or in the fibrous cap of a plaque containing high amounts of extracellular matrix components, but not in the lipid‐rich areas. In combination with high‐resolution MRI, Gadofluorine M might enable the detection of thin‐cap fibroatheromas, also named the vulnerable plaque. Copyright © 2007 John Wiley & Sons, Ltd.     

Utilization of nanoparticles as X‐ray contrast agents for diagnostic imaging applications

Among all the diagnostic imaging modalities, X‐ray imaging techniques are the most commonly used owing to their high resolution and low cost. The improvement of these techniques relies heavily on the development of novel X‐ray contrast agents, which are molecules that enhance the visibility of internal structures within the body in X‐ray imaging. To date, clinically used X‐ray contrast agents consist mainly of small iodinated molecules that might cause severe adverse effects (e.g. allergies, cardiovascular diseases and nephrotoxicity) in some patients owing to the large and repeated doses that are required to achieve good contrast. For this reason, there is an increasing interest in the development of alternative X‐ray contrast agents utilizing elements with high atomic numbers (e.g. gold, bismuth, ytterbium and tantalum), which are well known for exhibiting high absorption of X‐rays. Nanoparticles (NPs) made from these elements have been reported to have better imaging properties, longer blood circulation times and lower toxicity than conventional iodinated X‐ray contrast agents. Additionally, the combination of two or more of these elements into a single carrier allows for the development of multimodal and hybrid contrast agents. Herein, the limitations of iodinated X‐ray contrast agents are discussed and the parameters that influence the efficacy of X‐ray contrast agents are summarized. Several examples of the design and production of both iodinated and iodine‐free NP‐based X‐ray contrast agents are then provided, emphasizing the studies performed to evaluate their X‐ray attenuation capabilities and their toxicity in vitro and in vivo. Copyright © 2014 John Wiley & Sons, Ltd.     

Aerosols and gaseous contrast agents for magnetic resonance imaging of the lung

Magnetic resonance imaging of lungs and the investigation of pulmonary pathologies with this technique are limited by low proton spin density, degraded magnetic homogeneity and motion. Inhaled contrast agents (gases or aerosols) can improve the diagnostic value of MRI for lung. Paramagnetic contrast agents such as gadolinium chelates aerosol or dioxygen gas increase the relaxivity of proton in lung parenchyma and can be used to assess the ventilated fraction of the bronchoalveolar space. Similarly, inhalation of non proton‐MRI nuclei such as perfluorinated gas or hyperpolarized gases (³He or ¹²⁹Xe) can provide functional ventilation image. In this review paper, the principles, the practical implementation, the limitations and possible safety issues of these different techniques are summarized. The main pre‐clinical and clinical applications of these approaches based on oral contrast agents are reviewed and illustrated with cutting‐edge lung MRI studies. Copyright © 2008 John Wiley & Sons, Ltd.     

Functionalized single‐walled carbon nanotubes containing traces of iron as new negative MRI contrast agents for in vivo imaging

Single‐walled carbon nanotubes (SWCNTs) containing traces of iron oxide were functionalized by noncovalent lipid‐PEG or covalent carboxylic acid function to supply new efficient MRI contrast agents for in vitro and in vivo applications. Longitudinal (r₁) and transversal (r₂) water proton relaxivities were measured at 300 MHz, showing a stronger T₂ feature as an MRI contrast agent (r₂/r₁ = 190 for CO₂H functionalisation). The r₂ relaxivity was demonstrated to be correlated to the presence of iron oxide in the SWNT‐carboxylic function COOH, in comparison to iron‐free ones. Biodistribution studies on mice after a systemic injection showed a negative MRI contrast in liver, suggesting the presence of the nanotubes in this organ until 48 h after i.v. injection. The presence of carbon nanotubes in liver was confirmed after ex vivo carbon extraction. Finally, cytotoxicity studies showed no apparent effect owing to the presence of the carbon nanotubes. The functionalized carbon nanotubes were well tolerated by the animals at the dose of 10 µg g^?1 body weight. Copyright © 2012 John Wiley & Sons, Ltd.     

多模态超声造影剂的现状及研究进展

近年来,随着多种医学成像技术的相互融合和分子影像技术的迅速发展,以超声分子显像为基础,同时具有多种影像学对比显影能力的多模态超声造影剂已成为当前超声影像学领域的研究热点之一。多模态超声造影剂不仅可以提高超声成像的分辨力、敏感性、对比度,还能在一定程度改善传统单一影像诊断的局限性,实现多种方式成像的优势互补,具有广阔的应用前景。本文就多模态超声造影剂的现状及进展进行综述。     

Cellular uptake and imaging studies of gadolinium‐loaded single‐walled carbon nanotubes as MRI contrast agents

We quantify here, for the first time, the intracellular uptake (J774A.1 murine macrophage cells) of gadolinium‐loaded ultra‐short single‐walled carbon nanotubes (gadonanotubes or GNTs) in a 3 T MRI scanner using R₂ and R₂* mapping in vitro. GNT‐labeled cells exhibited high and linear changes in net transverse relaxations (ΔR₂ and ΔR) with increasing cell concentration. The measured ΔR₂* were about three to four times greater than the respective ΔR₂ for each cell concentration. The intracellular uptake of GNTs was validated with inductively coupled plasma optical emission spectrometry (ICP‐OES), indicating an average cellular uptake of 0.44 ± 0.09 pg Gd per cell or 1.69 × 10⁹ Gd³⁺ ions per cell. Cell proliferation MTS assays demonstrated that the cells were effectively labeled, without cytotoxicity, for GNTs concentrations ≤28 µM Gd. In vivo relaxometry of a subcutaneously‐injected GNT‐labeled cell pellet in a mouse was also demonstrated at 3 T. Finally, the pronounced R₂* effect of GNT‐labeled cells enabled successful in vitro visualization of labeled cells at 9.4 T. Copyright © 2010 John Wiley & Sons, Ltd.     

Clinical and biological consequences of transmetallation induced by contrast agents for magnetic resonance imaging: a review

Gadolinium-based contrast agents (CAs) are widely used to enhance the contrast of images in magnetic resonance imaging procedures. Two categories of gadolinium chelates exist: the macrocyclic molecules where Gd3+ is caged in the pre-organized cavity of the ligand and the linear molecules. Gadolinium chelates differ in their thermodynamic stability constants and in their kinetic stability. In general, macrocyclic chelates such as Gd-DOTA or Gd-HP-DO3A are more stable than linear molecules. Even among linear agents, differences can be found. There is increasing evidence that transmetallation can be found in vivo, in the case of certain CAs (especially linear chelates), with body cations such as zinc, calcium or iron. Furthermore, analytical interference with colorimetric determination of calcium has been clinically evidenced with two linear chelates, Gd-DTPA-BMA and Gd-DTPA-BMEA. Clinical cases of spurious hypocalcaemia have been reported with these molecules. Such interference with some colorimetric assays for calcium is clinically relevant in that it can lead to unnecessary and potentially harmful treatment for hypocalcaemia.     

Modulating the relaxivity of hyperpolarized substrates with gadolinium contrast agents

Gadolinium contrast agents are known to affect the outer‐sphere relaxivities of nuclei other than protons. We have investigated the heteronuclear relaxivities of four commercial gadolinium contrast agents (Magnevist, Omniscan, Dotarem and Gadovist) on the relaxation of ¹³C in glycine, ¹³C in pyruvate and ¹⁵N in choline with long relaxation times. Marked differences in the relaxivities were found between different contrast agents and are attributed to electrostatic effects. This methodology may find applications in the field of hyperpolarized magnetic resonance and by way of example we show that injection of a bolus of contrast agent into an aqueous solution containing hyperpolarized ¹⁵N labeled tetramethylammonium or ¹³C labeled pyruvate leads to a predictable shortening of the lifetime of the hyperpolarized signal. Copyright © 2009 John Wiley & Sons, Ltd.     

纳米级微泡:一种新的超声造影剂

随着分子生物学技术的发展和延伸,超声分子成像获得了飞速的发展,但由于常规造影剂微泡不能透过血管壁,超声分子成像的研究仅限于血管内的靶分子.最近纳米级微泡超声造影剂的出现为针对肿瘤细胞的超声分子成像带来了希望.本文对纳米级微泡超声造影剂的研究背景、研究现状及下一步的研究与应用进行了综述.     

新型含钆磁共振对比剂在小鼠体内的成像研究

目的 研究一种新型含钆磁共振对比剂（Gd-PPF-S-CAs）在小鼠体内正常器官的成像研究。 方法 首先将Gd-PPF-S-CAs与小分子磁共振对比剂（DTPA-Gd）在体外弛豫效能进行对比;其次选取正常Balb/C小鼠10只,随机分为两组,一组注射Gd-PPF-S-CAs,一组注射临床用DTPA-Gd,分别进行小鼠的肝、肾、膀胱扫描,将Gd-PPF-S-CAs组在体内增强效果、增强幅度以及强化持续时间3个方面与临床DTPA-Gd进行对比。 结果 体外弛豫效能结果显示,实验组Gd-PPF-S-CAs弛豫率为15.43 mmol^-1s^-1,是临床DTPA-Gd弛豫率（3.53 mmol^-1s^-1）的5倍;通过对小鼠肝、肾、膀胱的增强效果、增强幅度以及强化持续时间进行分析,Gd-PPF-S-CAs较临床用DTPA-Gd在小鼠肝、肾、膀胱有着更为明显的增强效果、较高的增强幅度以及长时间的强化持续窗口。 结论 Gd-PPF-S-CAs在体外有着较高的弛豫效能;在小鼠体内正常器官有着明显的增强效果和长效的强化持续时间,能够有效的解决小分子临床DTPA-Gd的增强幅度较低、组织对比度不高和成像窗口时间较短的不足;同时,Gd-PPF-S-CAs具有酶降解特性,能够在体内快速代谢,有效地解决了Gd³⁺对比剂的潜在毒性问题,具有良好的临床应用前景。     

A review of responsive MRI contrast agents: 2005–2014

This review focuses on MRI contrast agents that are responsive to a change in a physiological biomarker. The response mechanisms are dependent on six physicochemical characteristics, including the accessibility of water to the agent, tumbling time, proton exchange rate, electron spin state, MR frequency or superparamagnetism of the agent. These characteristics can be affected by changes in concentrations or activities of enzymes, proteins, nucleic acids, metabolites, or metal ions, or changes in redox state, pH, temperature, or light. A total of 117 examples are presented, including ones that employ nuclei other than ¹H, which attests to the creativity of multidisciplinary research efforts to develop responsive MRI contrast agents. Copyright © 2014 John Wiley & Sons, Ltd.     

超声分子影像学在疾病诊断方面的研究进展

随着分子影像学的出现,靶向性超声造影剂在分子影像的应用愈来愈受到人们的关注.通过主动性和被动性两种靶向机制,超声造影剂可在分子水平无创性显示炎性反应、血栓、肿瘤的血管形成等,并且分子生物学和超声医学的良好发展前景,将为超声分子影像学开辟更大的应用空间.     

超顺磁性氧化铁纳米粒子在脑磁共振成像中的应用

超顺磁性氧化铁纳米粒子的磁性及生物相容性,使其在生物医学多个领域的应用研究都逐渐发展起来。本文介绍磁共振成像（MRI）及脑功能磁共振成像（fMRI）基本原理。列举不同性能的磁性氧化铁粒子作为磁共振成像对比剂在脑科学应用中的研究进展。表面结合单克隆抗体、蛋白质、多肽、核苷酸分子或其它特殊聚合物的磁性氧化铁粒子具有吸收特异性（靶向性）,结合MRI可实现对脑部病变前期改变、药物输运及治疗的监测,对细胞、生物分子包括mRNA的成像及探测。经葡聚糖或聚乙二醇修饰的超顺磁性氧化铁纳米粒子血液半衰期较长,可作为对比剂用于脑fMRI成像。控制氧化铁纳米粒子的粒度及表面修饰物的物理化学性质、提高饱和磁化强度、借以接枝以各种靶向性的物质、开发具有荧光-磁性等多种性能的复合纳米粒子及掌握纳米粒子与生物分子、细胞、及生物组织之间的相互作用,则需要更深入的研究。     

Dual nano‐sized contrast agents in PET/MRI: a systematic review

Nowadays molecular imaging plays a vital role in achieving a successful targeted and personalized treatment. Hence, the approach of combining two or more medical imaging modalities was developed. The objective of this review is to systematically compare recent dual contrast agents in Positron Emission Tomography (PET)/Magnetic Resonance Imaging (MRI) and in some cases Single photon emission computed tomography (SPECT)/MRI in terms of some their characteristics, such as tumor uptake, and reticuloendothelial system uptake (especially liver) and their relaxivity rates for early detection of primary cancer tumor. To the best of our knowledge, this is the first systematic and integrated overview of this field. Two reviewers individually directed the systematic review search using PubMed, MEDLINE and Google Scholar. Two other reviewers directed quality assessment, using the criteria checklist from the CAMARADES (Collaborative Approach to Meta‐Analysis and Review of Animal Data from Experimental Studies) tool, and differences were resolved by consensus. After reviewing all 49 studies, we concluded that a size range of 20–200 nm can be used for molecular imaging, although it is better to try to achieve as small a size as it is possible. Also, small nanoparticles with a hydrophilic coating and positive charge are suitable as a T₂ contrast agent. According to our selected data, the most successful dual probes in terms of high targeting were with an average size of 40 nm, PEGylated using peptides as a biomarker and radiolabeled with copper 64 and gallium 68. Copyright © 2017 John Wiley & Sons, Ltd.     

Ultrasonic imaging of endothelial CD81 expression using CD81-targeted contrast agents in in vitro and in vivo studies

This study is designed to investigate the feasibility for molecular imaging of endothelial CD81 expression in vitro and in vivo using the CD81-targeted ultrasound contrast agents (UCA). In the in vitro study, murine bEnd.3 cells were stimulated with phenazine methosulfate (PMS), an oxidative stress inducer. Changes in CD81 expression after stimulation were confirmed by Western blotting, tracked by using the targeted UCA and further imaged under ultrasound imaging system with 5 MHz transmit frequency. In the in vivo study, expression of endothelial CD81 proteins in murine carotid artery vessels was studied using high-frequency ultrasound system with 40 MHz transmit frequency. Our results showed that endothelial CD81 expression was gradually up-regulated with the increase of PMS concentration. Correspondingly, the accumulation of targeted UCA was gradually improved and could be inhibited significantly upon addition of free anti-CD81 antibodies. The mean video intensity (grey-level) of stimulated cells and vessels from backscatter of the CD81-targeted UCA was 17.2 (interquartile range [IQR] 15.4–19.8) and 27.2 (IQR 22.4–29.8), significantly greater than that of non-stimulated cells with 9.0 (IQR 8.6–10.8) (p < 0.01) and non-stimulated vessels with 11.3 (IQR 10.4–13.2) (p < 0.01), respectively. In conclusion, CD81-targeted UCA allows noninvasive assessment of the expression levels of CD81 on the vascular endothelium and may provide potential insights into early atherosclerotic plaque detection and treatment monitoring.