磁共振造影剂的研究进展

随着磁共振影像技术的快速发展,MRI在医学领域得到广泛应用,已成为目前临床常规影像诊断方法和手段之一.但MRI对信号探测的敏感性较低,因此需要某些介质在靶组织内大量聚集以达到信号扩增的目的,于是磁共振成像对比剂应用而生.磁共振造影剂(对比剂)可以提高成像分辨率,增加正常与病变组织的成像对比度,从而提高磁共振诊断疾病的敏感性和特异性,目前逐日成为众多学者研究关注的焦点之一.超顺磁性氧化铁纳米粒是一种新型的磁共振对比剂,它的有效成份为纳米级的Fe3O4或Fe2O3晶体核心,主要通过缩短组织中成像水质子的弛豫时间从而加快组织弛豫速率,得以提高正常组织和病灶组织的成像信号对比度,对肝、脾、淋巴结病变的成像效果好,安全性高,能够显著提高小病灶的检出,从而达到早期诊断发现疾病的目的.本文主要就磁共振造影剂的原理、分类及研究进展,尤其是超顺磁性氧化铁在肝脏疾病诊断中的应用进行了综述,并且对磁共振造影剂的未来发展趋势进行了展望.     

磁共振间质淋巴造影实验研究

目的:探讨间质MR淋巴造影对肢体淋巴水肿的诊断价值。方法:用改良的Danese手术方法在13只新西兰大白兔后肢一侧形成淋巴水肿模型,另一侧作为对照。在每只大白兔双侧后肢足背部趾蹼处注射0.2ml欧乃影,按摩注射部位30秒钟。分别于造影剂注射前后进行三维MR淋巴造影及延迟淋巴造影成像。结果:实验侧淋巴管阻塞早期为渗出性改变及淋巴管侧支开放,晚期出现淋巴管扩张、迂曲、皮肤逆流。引流远端淋巴结显影较对照侧明显延迟。对照侧欧乃影吸收迅速,引流区域各组淋巴结、淋巴管及胸导管显示清晰。结论:间质MR淋巴造影可以在解剖背景下很好地显示引流区域淋巴管、淋巴结的解剖形态、功能及其异常表现。     

碳量子点的合成、表征及应用

对碳量子点的常用合成方法、表征手段及应用情况进行介绍。碳量子点是一种以碳元素为主体的新型荧光纳米材料,具有光学性能优良、细胞毒性低、生物相容性好、易于功能化和成本低廉等优点,在生物和医药领域具有广阔的开发前景。     

肢体淋巴水肿磁共振淋巴造影技术方法的实验研究

目的：研究肢体水肿演变过程中不同时期的胍淋巴造影影像特征夏其病理基础,探讨MR淋巴造影在肢体淋巴水肿方面的诊断价值。方法：用改良的Danese手术方法在20只新西兰大白兔后肢一侧形成淋巴水肿模型,另一侧作为对照。在每只大白兔双侧后肢足背部趾硅处注射0.2ml欧乃影,于淋巴水肿演变过程的不同时期进行三维胍淋巴造影。结果：胍淋巴造影能准确地确定淋巴管阻塞的部位,反映淋巴管形态、功能的状况。肢体淋巴水肿的不同时期,由于其病理基础不同。产生不同的MR淋巴造影表现。结论：间质MR淋巴造影可以在解剖背景下敏感而又可靠地显示各期肢体淋巴水肿。     

聚乙二醇丙烯酸酯的合成与表征

以聚乙二醇-400（PEG400）与丙烯酸直接缩合反应,在不加有毒带水剂的条件下合成了丙烯酸聚乙二醇酯（PEGA）。通过正交实验确定酯化反应的最佳条件：丙烯酸／PEG400的摩尔比为2．0：1．0,反应温度是110℃,阻聚剂对苯二酚为0．4％（以醇酸总质量计）,反应时间为6小时,催化剂对甲苯磺酸为0．8％（以醇酸总质量计）,产率为76．7％。产品结构经IR和1HNMR表征,证明是所需的产物。     

磁共振淋巴造影技术方法研究

目的：评估欧乃影介导的磁共振间质淋巴造影在诊断前列腺癌盆腔淋巴转移的可行性和安全性。方法：所有30例病理确诊前列腺癌患者,采用1.5T菲利浦超导型磁共振成像仪检查。先完成盆腔前列腺平扫,接着皮下注射造影剂欧乃影行三维增强磁共振淋巴造影术（3D CE-MRL）,最后欧乃影静脉注射盆腔前列腺增强扫描。结果：皮下注射后,欧乃影迅速吸收进入淋巴系统,引流区域各组淋巴结、淋巴管显示清晰。结论：皮下注射＂欧乃影＂行磁共振间质淋巴造影术诊断前列腺癌患者盆腔淋巴转移是可行和安全的。     

肢体淋巴水肿磁共振淋巴造影技术方法的实验研究

目的:研究肢体水肿演变过程中不同时期的MR淋巴造影影像特征及其病理基础,探讨MR淋巴造影在肢体淋巴水肿方面的诊断价值。方法:用改良的Danese手术方法在20只新西兰大白兔后肢一侧形成淋巴水肿模型,另一侧作为对照。在每只大白兔双侧后肢足背部趾蹼处注射0.2ml欧乃影,于淋巴水肿演变过程的不同时期进行三维MR淋巴造影。结果:MR淋巴造影能准确地确定淋巴管阻塞的部位,反映淋巴管形态、功能的状况。肢体淋巴水肿的不同时期,由于其病理基础不同,产生不同的MR淋巴造影表现。结论:间质MR淋巴造影可以在解剖背景下敏感而又可靠地显示各期肢体淋巴水肿。     

超顺磁性氧化铁微粒磁共振造影剂的制备研究

本文报道自制一种超顺磁性氧化铁微粒,探讨了各种制备条件对氧化铁颗粒大小、稳定性及弛豫增强性能的影响,分析了体外磁共振扫描时超顺磁性氧化铁的浓度——增强效果关系,初步提出了T_1W及T_2W成像时的最适造影浓度。     

阳离子聚丙烯酰胺的合成及其表征

合成了阳离子功能单体甲基丙烯酰氧乙基二甲基丁基溴化铵(DMB),并将DMB与丙烯酰胺(AM)共聚制备了阳离子聚丙烯酰胺,并用IR,NMR对其结构进行了表征。     

维生素E琥珀酸酯的合成及表征

目的:合成维生素E琥珀酸酯并对其进行表征.方法:以d,1-α-生育酚和丁二酸酐为原料,吡啶为溶媒,合成了维生素E琥珀酸酯.采用紫外光谱法、红外光谱法、核磁共振氢谱和差示扫描量热分析对产物进行表征.结果:经验证,成功合成了维生素E琥珀酸酯.结论:成功合成了维生素E琥珀酸酯.     

Preparation,Purification, and Characterization of Lanthanide Complexes for Use as Contrast Agents for Magnetic Resonance Imaging

Polyaminopolycarboxylate-based ligands are commonly used to chelate lanthanide ions, and the resulting complexes are useful as contrast agents for magnetic resonance imaging (MRI). Many commercially available ligands are especially useful because they contain functional groups that allow for fast, high-purity, and high-yielding conjugation to macromolecules and biomolecules via amine-reactive activated esters and isothiocyanate groups or thiol-reactive maleimides. While metalation of these ligands is considered common knowledge in the field of bioconjugation chemistry, subtle differences in metalation procedures must be taken into account when selecting metal starting materials. Furthermore, multiple options for purification and characterization exist, and selection of the most effective procedure partially depends on the selection of starting materials. These subtle differences are often neglected in published protocols. Here, our goal is to demonstrate common methods for metalation, purification, and characterization of lanthanide complexes that can be used as contrast agents for MRI (Figure 1). We expect that this publication will enable biomedical scientists to incorporate lanthanide complexation reactions into their repertoire of commonly used reactions by easing the selection of starting materials and purification methods.     

New Dual Mode Gadolinium Nanoparticle Contrast Agent for Magnetic Resonance Imaging

Background

Liposomal-based gadolinium (Gd) nanoparticles have elicited significant interest for use as blood pool and molecular magnetic resonance imaging (MRI) contrast agents. Previous generations of liposomal MR agents contained gadolinium-chelates either within the interior of liposomes (core-encapsulated gadolinium liposomes) or presented on the surface of liposomes (surface-conjugated gadolinium liposomes). We hypothesized that a liposomal agent that contained both core-encapsulated gadolinium and surface-conjugated gadolinium, defined herein as dual-mode gadolinium (Dual-Gd) liposomes, would result in a significant improvement in nanoparticle-based T1 relaxivity over the previous generations of liposomal agents. In this study, we have developed and tested, both in vitro and in vivo, such a dual-mode liposomal-based gadolinium contrast agent.

Methodology/Principal Findings

Three types of liposomal agents were fabricated: core-encapsulated, surface-conjugated and dual-mode gadolinium liposomes. In vitro physico-chemical characterizations of the agents were performed to determine particle size and elemental composition. Gadolinium-based and nanoparticle-based T1 relaxivities of various agents were determined in bovine plasma. Subsequently, the agents were tested in vivo for contrast-enhanced magnetic resonance angiography (CE-MRA) studies. Characterization of the agents demonstrated the highest gadolinium atoms per nanoparticle for Dual-Gd liposomes. In vitro, surface-conjugated gadolinium liposomes demonstrated the highest T1 relaxivity on a gadolinium-basis. However, Dual-Gd liposomes demonstrated the highest T1 relaxivity on a nanoparticle-basis. In vivo, Dual-Gd liposomes resulted in the highest signal-to-noise ratio (SNR) and contrast-to-noise ratio in CE-MRA studies.

Conclusions/Significance

The dual-mode gadolinium liposomal contrast agent demonstrated higher particle-based T1 relaxivity, both in vitro and in vivo, compared to either the core-encapsulated or the surface-conjugated liposomal agent. The dual-mode gadolinium liposomes could enable reduced particle dose for use in CE-MRA and increased contrast sensitivity for use in molecular imaging.     

Development and Characterization of an Antibody-Labeled Super-Paramagnetic Iron Oxide Contrast Agent Targeting Prostate Cancer Cells for Magnetic Resonance Imaging

In this study we developed, characterized and validated in vitro a functional superparagmagnetic iron-oxide based magnetic resonance contrast agent by conjugating a commercially available iron oxide nanoparticle, Molday ION Rhodamine-B Carboxyl (MIRB), with a deimmunized mouse monoclonal antibody (muJ591) targeting prostate-specific membrane antigen (PSMA). This functional contrast agent is intended for the specific and non-invasive detection of prostate cancer cells that are PSMA positive, a marker implicated in prostate tumor progression and metastasis. The two-step carbodiimide reaction used to conjugate the antibody to the nanoparticle was efficient and we obtained an elemental iron content of 1958±611 per antibody. Immunofluorescence microscopy and flow cytometry showed that the conjugated muJ591:MIRB complex specifically binds to PSMA-positive (LNCaP) cells. The muJ591:MIRB complex reduced cell adhesion and cell proliferation on LNCaP cells and caused apoptosis as tested by Annexin V assay, suggesting anti-tumorigenic characteristics. Measurements of the T2 relaxation time of the muJ591:MIRB complex using a 400 MHz Innova NMR and a multi-echo spin-echo sequence on a 3T MRI (Achieva, Philips) showed a significant T2 relaxation time reduction for the muJ591:MIRB complex, with a reduced T2 relaxation time as a function of the iron concentration. PSMA-positive cells treated with muJ591:MIRB showed a significantly shorter T2 relaxation time as obtained using a 3T MRI scanner. The reduction in T2 relaxation time for muJ591:MIRB, combined with its specificity against PSMA+LNCaP cells, suggest its potential as a biologically-specific MR contrast agent.     

Imaging Lymphatic System in Breast Cancer Patients with Magnetic Resonance Lymphangiography

Objective

To investigate the feasibility of gadolinium (Gd) contrast-enhanced magnetic resonance lymphangiography (MRL) in breast cancer patients within a typical clinical setting, and to establish a Gd-MRL protocol and identify potential MRL biomarkers for differentiating metastatic from non-metastatic lymph nodes.

Materials and Methods

32 patients with unilateral breast cancer were enrolled and divided into 4 groups of 8 patients. Groups I, II, and III received 1.0, 0.5, and 0.3 ml of intradermal contrast; group IV received two 0.5 ml doses of intradermal contrast. MRL images were acquired on a 3.0 T system and evaluated independently by two radiologists for the number and size of enhancing lymph nodes, lymph node contrast uptake kinetics, lymph vessel size, and contrast enhancement patterns within lymph nodes.

Results

Group III patients had a statistically significant decrease in the total number of enhancing axillary lymph nodes and lymphatic vessels compared to all other groups. While group IV patients had a statistically significant faster time to reach the maximum peak enhancement over group I and II (by 3 minutes), there was no other statistically significant difference between imaging results between groups I, II, and IV. 27 out of 128 lymphatic vessels (21%) showed dilatation, and all patients with dilated lymphatic vessels were pathologically proven to have metastases. Using the pattern of enhancement defects as the sole criterion for identifying metastatic lymph nodes during Gd-MRL interpretation, and using histopathology as the gold standard, the sensitivity and specificity were estimated to be 86% and 95%, respectively.

Conclusion

Gd-MRL can adequately depict the lymphatic system, can define sentinel lymph nodes, and has the potential to differentiate between metastatic and non-metastatic lymph nodes in breast cancer patients.     

Pathological Mechanism for Delayed Hyperenhancement of Chronic Scarred Myocardium in Contrast Agent Enhanced Magnetic Resonance Imaging

Objectives

To evaluate possible mechanism for delayed hyperenhancement of scarred myocardium by investigating the relationship of contrast agent (CA) first pass and delayed enhancement patterns with histopathological changes.

Materials and Methods

Eighteen pigs underwent 4 weeks ligation of 1 or 2 diagonal coronary arteries to induce chronic infarction. The hearts were then removed and perfused in a Langendorff apparatus. The hearts firstly experienced phosphorus 31 MR spectroscopy. The hearts in group I (n = 9) and II (n = 9) then received the bolus injection of Gadolinium diethylenetriamine pentaacetic acid (0.05 mmol/kg) and gadolinium-based macromolecular agent (P792, 15 µmol/kg), respectively. First pass T₂ ^* MRI was acquired using a gradient echo sequence. Delayed enhanced T₁ MRI was acquired with an inversion recovery sequence. Masson''s trichrome and anti- von Willebrand Factor (vWF) staining were performed for infarct characterization.

Results

Wash-in of both kinds of CA caused the sharp and dramatic T₂ ^* signal decrease of scarred myocardium similar to that of normal myocardium. Myocardial blood flow and microvessel density were significantly recovered in 4-week-old scar tissue. Steady state distribution volume (ΔR₁ relaxation rate) of Gd-DTPA was markedly higher in scarred myocardium than in normal myocardium, whereas ΔR₁ relaxation rate of P792 did not differ significantly between scarred and normal myocardium. The ratio of extracellular volume to the total water volume was significantly greater in scarred myocardium than in normal myocardium. Scarred myocardium contained massive residual capillaries and dilated vessels. Histological stains indicated the extensively discrete matrix deposition and lack of cellular structure in scarred myocardium.

Conclusions

Collateral circulation formation and residual vessel effectively delivered CA into scarred myocardium. However, residual vessel without abnormal hyperpermeability allowed Gd-DTPA rather than P792 to penetrate into extravascular compartment. Discrete collagen fiber meshwork and loss of cellularity enlarged extracellular space accessible to Gd-DTPA, resulting in the delayed hyper-enhanced scar.     

Modeling the Effect of Intra-Voxel Diffusion of Contrast Agent on the Quantitative Analysis of Dynamic Contrast Enhanced Magnetic Resonance Imaging

Quantitative dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) provides estimates of physiologically relevant parameters related to tissue blood flow, vascular permeability, and tissue volume fractions which can then be used for prognostic and diagnostic reasons. However, standard techniques for DCE-MRI analysis ignore intra-voxel diffusion, which may play an important role in contrast agent distribution and voxel signal intensity and, thus, will affect quantification of the aforementioned parameters. To investigate the effect of intra-voxel diffusion on quantitative DCE-MRI, we developed a finite element model of contrast enhancement at the voxel level. For diffusion in the range of that expected for gadolinium chelates in tissue (i.e., 1×10⁻⁴ to 4×10⁻⁴ mm²/s), parameterization errors range from −58% to 12% for K^trans, −9% to 8% for v_e, and −60% to 213% for v_p over the range of K^trans, v_e, v_p, and temporal resolutions investigated. Thus the results show that diffusion has a significant effect on parameterization using standard techniques.     

Novel Gd Nanoparticles Enhance Vascular Contrast for High-Resolution Magnetic Resonance Imaging

Background

Gadolinium (Gd), with its 7 unpaired electrons in 4f orbitals that provide a very large magnetic moment, is proven to be among the best agents for contrast enhanced MRI. Unfortunately, the most potent MR contrast agent based on Gd requires relatively high doses of Gd. The Gd-chelated to diethylene-triamine-penta-acetic acid (DTPA), or other derivatives (at 0.1 mmole/kg recommended dose), distribute broadly into tissues and clear through the kidney. These contrast agents carry the risk of Nephrogenic Systemic Fibrosis (NSF), particularly in kidney impaired subjects. Thus, Gd contrast agents that produce higher resolution images using a much lower Gd dose could address both imaging sensitivity and Gd safety.

Methodology/Principal Findings

To determine whether a biocompatible lipid nanoparticle with surface bound Gd can improve MRI contrast sensitivity, we constructed Gd-lipid nanoparticles (Gd-LNP) containing lipid bound DTPA and Gd. The Gd-LNP were intravenously administered to rats and MR images collected. We found that Gd in Gd-LNP produced a greater than 33-fold higher longitudinal (T₁) relaxivity, r₁, constant than the current FDA approved Gd-chelated contrast agents. Intravenous administration of these Gd-LNP at only 3% of the recommended clinical Gd dose produced MRI signal-to-noise ratios of greater than 300 in all vasculatures. Unlike current Gd contrast agents, these Gd-LNP stably retained Gd in normal vasculature, and are eliminated predominately through the biliary, instead of the renal system. Gd-LNP did not appear to accumulate in the liver or kidney, and was eliminated completely within 24 hrs.

Conclusions/Significance

The novel Gd-nanoparticles provide high quality contrast enhanced vascular MRI at 97% reduced dose of Gd and do not rely on renal clearance. This new agent is likely to be suitable for patients exhibiting varying degrees of renal impairment. The simple and adaptive nanoparticle design could accommodate ligand or receptor coating for drug delivery optimization and in vivo drug-target definition in system biology profiling, increasing the margin of safety in treatment of cancers and other diseases.     

In Vitro Longitudinal Relaxivity Profile of Gd(ABE-DTTA), an Investigational Magnetic Resonance Imaging Contrast Agent

Purpose

MRI contrast agents (CA) whose contrast enhancement remains relatively high even at the higher end of the magnetic field strength range would be desirable. The purpose of this work was to demonstrate such a desired magnetic field dependency of the longitudinal relaxivity for an experimental MRI CA, Gd(ABE-DTTA).

Materials and Methods

The relaxivity of 0.5mM and 1mM Gd(ABE-DTTA) was measured by Nuclear Magnetic Relaxation Dispersion (NMRD) in the range of 0.0002 to 1T. Two MRI and five NMR instruments were used to cover the range between 1.5 to 20T. Parallel measurement of a Gd-DTPA sample was performed throughout as reference. All measurements were carried out at 37°C and pH 7.4.

Results

The relaxivity values of 0.5mM and 1mM Gd(ABE-DTTA) measured at 1.5, 3, and 7T, within the presently clinically relevant magnetic field range, were 15.3, 11.8, 12.4 s^-1mM^-1 and 18.1, 16.7, and 13.5 s^-1mM^-1, respectively. The control 4 mM Gd-DTPA relaxivities at the same magnetic fields were 3.6, 3.3, and 3.0 s^-1mM^-1, respectively.

Conclusions

The longitudinal relaxivity of Gd(ABE-DTTA) measured within the presently clinically relevant field range is three to five times higher than that of most commercially available agents. Thus, Gd(ABE-DTTA) could be a practical choice at any field strength currently used in clinical imaging including those at the higher end.     

In vivo Macrophage Imaging Using MR Targeted Contrast Agent for Longitudinal Evaluation of Septic Arthritis

Macrophages are key-cells in the initiation, the development and the regulation of the inflammatory response to bacterial infection. Macrophages are intensively and increasingly recruited in septic joints from the early phases of infection and the infiltration is supposed to regress once efficient removal of the pathogens is obtained. The ability to identify in vivo macrophage activity in an infected joint can therefore provide two main applications: early detection of acute synovitis and monitoring of therapy.In vivo noninvasive detection of macrophages can be performed with magnetic resonance imaging using iron nanoparticles such as ultrasmall superparamagnetic iron oxide (USPIO). After intravascular or intraarticular administration, USPIO are specifically phagocytized by activated macrophages, and, due to their magnetic properties, induce signal changes in tissues presenting macrophage infiltration. A quantitative evaluation of the infiltrate is feasible, as the area with signal loss (number of dark pixels) observed on gradient echo MR images after particles injection is correlated with the amount of iron within the tissue and therefore reflects the number of USPIO-loaded cells.We present here a protocol to perform macrophage imaging using USPIO-enhanced MR imaging in an animal model of septic arthritis, allowing an initial and longitudinal in vivo noninvasive evaluation of macrophages infiltration and an assessment of therapy action.