摘要
随着外科技术的迅速发展，人们要求影像学能检出更小的病变，更准确地定性，使得磁共振造影剂的研究备受关注。SPIO(超顺磁氧化铁)作为目前磁共振造影剂的研究热点，具有肝脏特异性，即主要由肝脏正常组织中的枯否氏细胞吞噬，而肿瘤组织由于缺乏该类细胞，使得SPIO在肿瘤部位含量低，从而可通过磁共振检查将正常与病变组织区分出来。SPIO在肝脏局灶性病变尤其是小肝癌、微小肝癌的检出率、定性鉴别诊断方面，开创了细胞功能与影像结合的新天地，使得肝脏疾病诊断程序变得更为简单、迅捷。此外，USPIO(超细顺磁氧化铁)造影剂在受体等介导的磁标记探针成像的应用前景也引起人们的广泛注意。所谓磁标记探针是指将顺磁性粒子以某种方式与特异性受体、抗体或基因结合转入细胞内，由于该顺磁性物质在细胞内的蓄积与所连接的物质数量成正比，因此，顺磁性物质引起的MR信号分布代表了受体、抗体或基因的分布，而通过顺磁性标记的配体，在受体介导下，产生特异性浓集，达到选择性强化进而成像的目的。我们的目的是通过详细考察各种因素对形成稳定的纳米Fe_3O_4磁流体的影响，设计合适的制备方案，制备出适合粒径范围的SPIO或USPIO，在活体及细胞水平检验其造影增强效果，并制订相应的质量标准和制各工艺，为工业化生产提供技术支持。
方法与内容：
With the rapid development of surgery, we want to get the better result from MR imaging, for example to find the disease with less area, and more accurately. It is more and more important to do some research on MR imaging contrast agents. The SPIO (superparamagnetic iron oxide) MR contrast agents have taken an important part in this field. This is partly because of its liver specification. The main part of SPIO is licked up by Kuffe's cells in normal liver. The tumor didn't contain these cells so SPIO concentration was lower in it. The difference between the normal and tumor can be detected in MR imaging. It brought about the new develop of diagnosis in combination cell fuction with MR imaging in liver limit cancer such as little and micro tumors. It was easier and rapider for the diagnosis procedure in liver disease. Besides this, USPIO (Ultrasmall Superparamagnetic Iron Oxide) contrast agents had a very potent future in so called magnetic labeled probe MR imaging. This probe would combine in some way with specific receptor, antibody and gene and go into the cell. The concentration of the magnetic cores inside the cell correlate with the material they could bind with. So the MR imaging signal represent the distribution of the receipt, antibody and gene. What we want to is to study several important factors in
引文
1 詹松华，吴沛宏，杨振燕，主编．MRI．临床医师必读．科学出版社．2003；15-18
2 许乙凯，主编．磁共振造影剂及临床应用．人民卫生出版社．2003：16-56
3 Hapgspiel KD, Neidl KF, Eichenberger AC, et al. Detection of liver metastases: Coparision of superparamagnetic iron oxide-enchanced and unenchanced MR imaging at 1.5T with dynamic CT, intraoperative US, and percutaneous US. Radiology, 1995, 196(2): 471-478.
4 刘迎春，许妙琼，朱文祥．医疗诊断新技术—磁共振成像术．化学教育，2005(5)：4-7
5 徐隽，宋彬，闵鹏秋．肝脏磁共振对比剂研究现状及发展方向．临床放射学杂志，2001，20(9)：716-719
6 杨汉丰，周翔平．菲立磁增强肝脏磁共振成像．川北医学院学报，2004，19(3)：204-207
7 许乙凯．磁共振对比剂的发展概况及存在问题．第一军医大学学报，2002，22(9)：769-771
8 Saeed M, Wendland MF, Higgins CB. Blood-pool MR contrast agents for cardiovascular imaging. J Magn Reson Imaging. 2000, 12(6): 890-898.
9 Tombach B, Reimer P, Bremer C, Allkemper T, et al. First-pass and equilibrium-MRA of the aortoiliac region with a superparamagnetic iron oxide blood pool MR contrast agent (SHU 555 C): results of a human pilot study. NMR Biomed. 2004, 17(7): 500-506.
10 丁广良，胡红兵，卢广，等．两种新型的MRI造影剂．化学物理学报，2000，13(1)：99-102
11 Greco A, Mc Namara MT, Lan Thiez P, et al. Gadodiamide injection: nonionic gadolinium chelate for MR imaging of the brain and spine-phase Ⅱ-Ⅲ clinical trial. Radiology, 1990, 176(2): 451-456
12 Kabalka GW, Davis MA, Moss TH, et al. Gadolinium-labeled liposomes: targeted MR contrast agents for the liver and spleen. Radiology, 1987, 163(2): 255-258
13 Kato T, Suto Y, Matsuo T. Chondroitin sulfate iron colloid as an MR contrast agent for the hepatic reticuloendothelial system. J Comput Assist tomogr, 1993, 17(4): 603-608
14 Okuhata Y, Delivery of diagnostic agents for magnetic resonance imaging. Advanced Drug Delivery review, 1999, 37(1-3): 121-127
15 Helmberger T, Semelka RC. New contrast agents for imaging the liver. Magn Reson Imaging Clin N Am. 2001, 9(4): 745-766
16 Bonnemain B. Superparamagnetic agents in magnetic resonance imaging: physicochemical characteristics and clinical applications. A review. J Drug Target. 1998, 6(3): 167-174.
17 林红霞，陈骐．磁共振成像造影剂的研究进展．沈阳药科大学学报，2002，19(2)：138-142
18 Matsumura M, Yamada K, Fujimaki M, et al. Proton relaxation caused by Magnetic resonance imaging contrastagent, Oral Magnetic particles. Chem Pharm Bull, 1999, 47(6): 727-731.
19 Chu W, Paula J. Physical and chemical properties of suerparamagnetic iron oxide MR contrast agents: Ferumoxides, Ferumoxtran, Ferumoxsil. Mag Res Imaging, 1995, 13(5): 661-674.
20 Reimer P, Weissleder R, Lee AS, et al. Receptor imaging: application to MR imaging of liver cancer. Radiology, 1990, 177(3): 729-734.
21 Hamm B, Staks T, Taupitz M. Contrast-enhanced MR imaging of liver and spleen, first experience ihmans with new superparamagnetic iron oxide. J Magn Reson Imaging, 1994, 4(5): 659-668.
22 Dodd SJ, Williams M, Suhan JP, et al. Detection of single mammalian cells by high-resolution magnetic resonance imaging. Biophys J. 1999 Jan, 76(1): 103-109
23 Takahama K, Amano Y, Hayashi H, et al. T1-weighted magnetic resonance imaging sequence appropriate for the evaluation of the longitudinal relaxation effect of superparamagnetic iron oxide: a phantom study. J Nippon Med Sch. 2002, 69(6): 571-576.
24 Wang YX, Hussain SM, Krestin GP. Superparamagnetic iron oxide contrast agents: physicochemical characteristics and applications in MR imaging. Eur Radiol. 2001; 11(11): 2319-2331.
25 徐亮，郭启勇．超顺磁性氧化铁的研究现状及其在肝增强磁共振成像的临床应用及前景．国外医学临床放射学分册，2000，23(1)：31-36
26 刘岘．超顺磁性氧化铁类对比剂及其在肝病变的应用．国外医学临床放射学分册，2000，23(6)：325-327
27 Vogl TJ, Hammerstingl R, Schwarz W, et al. Superparamagnetic iron oxide-enhanced versus gadolinium-enhanced MR imaging for differental diagnosis of focal liver lesions, Radiology, 1996, 198: 881-893
28 Ward J, Naik KS, Guthrie JA, et al. Hepatic lesion detection: comparison of MRI after the administration of superparamagnetic iron oxide with dual phase CT by using alternative-free response receiver operating characteristic analysis. Radiology, 1999, 210(2): 459-466.
29 Weinmann HJ, Ebert W, Misselwitz B, et al. Tissue-specific MR contrast agents. Eur J Radiol. 2003, 46(1): 33-44.
30 Taupitz M, Schmitz S, Hamm B. Superparamagnetic iron oxide particles: current state and future development. Rofo Fortschr Geb Rontgenstr Neuen Bildgeb Verfahr. 2003, 175(6): 752-65.
31 Andreas EK, Michael L, Florian D, et al. MR Imaging of the liver with Resovist: safety, efficacy, and pharmacodynamic properties. Radiology, 1997, 204(3): 749-756
32 Wang YX, Hussian SM, Krestin GP. Superparamagnetic iron oxide contrast agents: physicochemical chacteristics and applications in MR imaging. Eur.Radiol, 2001,11:2319-2331
33 Sun R, Dittrich J, Le-Huu M, et al. Physical and Biological Characterization of Superparamagnetic Iron Oxide- and Ultrasmall Superparamagnetic Iron Oxide-Labeled Cells: A Comparison. Invest Radiol. 2005, 40(8): 504-513.
34 Varallyay P, Nesbit G, Muldoon LL, et al Comparison of two superparamagnetic viral-sized iron oxide particles ferumoxides and ferumoxtran-10 with a gadolinium chelate in imaging intracranial tumors. AJNR Am J Neuroradiol. 2002, 23(4):510-519.
35 Van Beers B E, Gallez B, Pringot J, et al. Contrast-enhanced MR imaging of the liver. Radiology, 1997, 203(1): 297-303
36 Harisinghai M G, Saini S, Slater GJ. et al. MR imaging of pelvic lymph nodes in primary pelvic carcinoma with ultrasmall superparamagnetic iron oxide (Combidex): Preliminary observations. JMRI, 1997, 7:161-169
37 Anzai Y, Prince MR. Iron oxide-enhanced MR lymphography: the evaluation of cervical lymph node metastases in head and neck cancer. J Magn Reson Imaging.1997, 7(1): 75-81.
38 Reimer P, Tombach B. Hepatic MRI with SPIO: detection and characterization of focal liver lesions. Eur Radiol, 1998;8(7):1198-1204.
39 Schmitz S A, Albrecht T, Wolf K-J. et al. MR angiography with superparamagnetic iron oxide: feasibility study. Radiology, 1999, 213(2): 603-611
40 Berry 1, Benderbous S, Ranjeva J P, et al. Contribution of Sinerem used as blood-pool contrast agent: detection of cerebral blood volume changes during apnea in the rabbit. MRM, 1996, 36: 415-423
41 Moore A, Basilion J P, Chiocca EA, et al. Measuring transferrin receptor gene expression by NMR imaging. Biochim Biop hys Acta, 1998, 1402(3): 239-249
42 张玉忠，张雪林．转铁蛋白受体与磁共振分子影像学研究近况．国外医学临床放射学分册，2004，27(5)：265-269
43 柴青芬，许乙凯，黄其鎏．磁共振靶向对比剂对研究进展．国外医学临床放射学分册，2000，23(5)：286-290
44 Weissleder R, Lee AS, Khaw BA, Shen T, et al. Antimyosin-labeled monocrystalline iron oxide allows detection of myocardial infarct: MR antibody imaging. Radiology, 1992, 182: 381-385
45 Weissleder R, Lee AS, Fischman AJ, et al. Polyclonal human immunoglobulin G labeled with polymeric iron oxide: antibody MR imaging. Radiology, 1991, 181: 245-249
46 Shaharabany M, Abramovitch R, Kushnir T, et al. In vivo molecular imaging of met tyrosine kinase growth factor receptor activity in normal organs and breast tumors. Cancer Res, 2001, 61: 4873～4881
47 何国祥，王毅翔．造影剂药理学及临床应用[M]．上海：上海科学技术出版社，2002．
48 王毛兰，胡春华，罗新．磁流体的制备、性质及其应用[J]．化学通报，2004，67(5)：31-37
49 Berkovdky BM, Medvedev VF, Krakov MS. Magnetic fluids-Engineering Applications. London: Oxford University Press, 1993
50 陈江．抗癌药物载体——磁流体的制备及其性质研究．中国医院药学杂志，1997，17(4)：163—165．
51 T. Kawaguchi, M. Hasegawa. Structure of dextran-magnetite complex: relation between conformation of dextran chains covering core and its molecular weight. J Mater Sci Mater Med. 2000, 11(1): 31-35
52 Groman EV, Josephson L, Lewis JM, et al. Biologically degradable superparamagnetic materials for use in clinical applications. 1989, US patent 4827945, section 7.1
53 Palmacci S, Josephson L. Synthesis of poiysaccharide covered superparamagnetic oxid colloids. 1993, US patent 5262176, example1
54 Groman EV, Josephson L, Lewis JM, at al. Biodegradable superparamagnetic matal oxides as contrast agents for MR imaging. 1990, US patent 4951675, section 6
55 Stillman AE, Wilke N, Li D, et al. Ultrasmall superparamagnetic iron oxide to enhance MRA of the renal and coronary arteries: studies in human patients. J Comput assist Tomogr, 1996, 20: 51-55
56 Stillman AE, Wilke N, Jerosch-Herold M. Use of anintravascular T contrast agent to improve MR cine myocardial-blood pool definition in man. J Magn Reson Imaging, 1997, 7: 765-767
57 Amano y, Herfkens RJ, Shifrin RY. et al. Three-dimensional cardiac cine magnetic resonance imaging with an ultrasmall superparamagnetic iron oxide blood pool agent (NC100150). J Magn Reson Imaging, 2000, 11(2): 81-86
58 Taylor AM, Panting JR, Keegan J, et al. Safety and preliminary findings with the intravascular contrast agent NC100150 injection for MR coronary angiography. J Magn Reson Imaging, 1999, 9(2): 220-227
59 陆敏杰，赵世华．超顺磁氧化铁在心血管MRI中的应用．国外医学临床放射学分册，2005，28(4)：266-269
60 Forsting M, Reith W, Dorfler A, et al. MRI in acute cerebral ischaemia: perfusion imaging with superparamagnetic iron oxide in a rat model. Neuroradiology, 1994, 36: 23-26
61 Simonsen CZ, Ostergaard l, Vestergaard-Poulsen P, et al. CBF and CBV measurements by USPIO bolus tracking: reproducibility and comparison with Gd-based values. Magn Reson Imaging, 1999, 9(3): 342-347
62 Chapon C, Franconi F, Lemaire L, et al. High field magnetic resonance imaging evaluation of superparamagnetic iron oxide nanoparticles in a permanent rat myocardial infarction. Invest Radiol, 2003, 38(3): 141-146
63 Ichikawa T, Arbab AS, Araki T, et al. Perfusion MR imaging with a superparamagnetic iron oxide using T_2-weighted and susceptibility-sensitive echoplanar sequences: evaluation of tumor vascularity in hepatocellular carcinoma. Am J Roentgenol, 1999, 173: 207-213.
64 Shen T, Weissleder R, Papisov M, et al. Monocrystalline iron oxide nanocompounds (MION): physicochemical properties. Magn Reson Med, 1993, 29(5): 599-601
65 Bulte JW, Zhang S, van-Gelderen P, et al. Neurotransplantation of magnetically labeled oligodendrocyte progenitors: magnetic resonance tracking of cell migration and myelination. Proc Natl Acad Sci USA, 1999, 96(26): 15256-15261
66 Moore A, Josephson L, Bhorade RM, et al. Human transferrin receptor gene as a marker gene for MR imaging. Radiology, 2001, 221(1): 244-250
67 Weissleder R, Reimer P, Lee AS, et al. MR receptor imaging: ultrasmall iron oxide particles targeted to asialoglycoprotein receptors. Am J Roentgenol, 1990, 155(6): 1161-1167
68 Weissleder R, Moore A, Mahmood U, et al. In vivo magnetic resonance imaging of transgene expression. Nat Med, 2000, 6(3): 351-355
69 Bremer C, Weissleder R. In vivo imaging of gene expression: MR and optical technologies. Acad Radiol, 2001, 8: 15～21
70 林曰增，张雪林．分子影像学研究进展．临床放射学杂志．2003，(22)1：77-80
71 许乙凯．磁共振对比剂的发展概况及存在问题．第一军医大学学报，2002，22(9)：769-771
72 吴传斌，高文伟，魏树礼．药用磁流体的研究和应用。军事医学院院刊，1994，18(1)：36-39
73 常津．医用磁性纳米材料的制备——制备条件对有效粒径的影响．中国生物医学工程学报，1996，15(4)：378-381
74 任鱼欢，刘勇健，牛亚丰．醇—水共热法制备Fe_3O_4磁流体．化工进展，2003，(1)：49-51
75 任鱼欢，庄虹，刘勇健．Fe_3O_4纳米颗粒的表面改性．化学研究，2003，14(1)：11-13
76 许乙凯，刘杏元，许璇，等．超顺磁性氧化铁粒子(SPIO)的弛豫率及磁化率研究．中国医学物理学杂志，1998，15(1)：9-10
77 王雨青，李铁福，马红岩，等．磁共振造影剂的处方筛选和稳定性研究．沈阳药科大学学报，2002，19(6)：395-397
78 许乙凯，陈永鹏，张嘉宁，等．肝细胞去唾液酸糖蛋白受体介导的超顺磁性氧化铁粒子磁共振增强研究．放射学实践，1999，14(3)：152-154
79 Lambert G, Fattal E, Couvreur P. Nanoparticulate systems for the delivery of antisense oligonucleotides. Adv Drug Deliv Rev, 2001, 47(1): 99-112
80 曹正国，周四维，孙凯，等．超顺磁性葡聚糖氧化铁纳米颗粒的制备及其作为基因载体的可行性研究．癌症，2004，23(10)：1105-1109
81 沃尔夫冈，肯尼斯，丹尼斯，等．造影剂．1999，CN1212629
82 王亭杰，王伟林，金涌，等．针状羟基氧化铁晶体的成核与生长．仪器仪表学报，1996，17(1)：399-403
83 阎玺庆，陈建海，蒋青峰，等．靶向药物载体——磁流体的制备及表面改性．第一军医大学学报，2005，25(6)：626-629
84 Kawaguchi. T, Hasegawa. T, Hasegawa. S, et, al. Dextran-magnetite complex: conformation of dextran chains and stability of solution. J Mat Sci: materials in Medicine, 2001, (12): 121-127
85 Standard infrared grating spectra volume 29-30, 28001-30000
86 Wessleder R, Stark DD, Englestad BL, et al. Superparamagnetic iron oxide: pharmacokinetics and toxicity. A JR, 1989, 152(1): 167-173
87 Guidance for industry: single dose acute toxicity testing for pharmaceuticals1 CEDR, 1996. 1～2
88 Toxicokinetics: the assessment of systemic exposure in toxicity studies1 CEDR, 1995. 9～10
89 Kopp AF, Laniado M, Dammann F, et al. MR imaging of the liver with Resovist: safety, efficacy, and pharmacodynamic properties. Radiology, 1997, 204: 749-756.
90 Kumano S, Murakami T, Kim T, et al. Using superparamagnetic iron oxide-enhanced MRI to differentiate metastatic hepatic tumors and nonsolid benign lesions. AJR Am J Roentgenol. 2003, 181(5): 1335-1339
91 Tanaka M, Nakashima O, Wada Y, et al. Pathomorphological study of Kupffer cells in hepatocellular carcinoma and hyperplastic nodular lesions in the liver. Hepatology. 1996, 24(4): 807-812
92 Clement O, Frija G, Chambon C, et al. Superparamagnetic iron oxide-enhanced magnetic resonance imaging of experimental liver tumors after mitomycin C administration. Invest Radiol. 1992, 27(3): 230-235
93 Winter TC, Freeny PC, Nghiem HV, et al. MR imaging with i.v. superparamagnetic iron oxide: efficacy in the detection of focal hepatic lesions. AJR Am J Roentgenol. 1993, 161(6): 1191-1198
94 邵成伟，王培军，田建明，等．直接注射法制作大鼠肝癌模型．中国医学影像技术，2002，18(8)：731-732
95 王锦波，何振平．移植法建立大鼠肝癌模型．实验动物科学与管理．1999，16(2)：26-27
96 李琦，孟庆莉，彭永海，等．大鼠移植性肝癌模型的制作．第二军医大学学报 2002，23(10)：1074
97 刘爱连，郎志谨，付维利，等．SD大鼠肝癌模型的MRI征象与病理对照研究． 中国医学计算机成像杂志，1999，5(1)：33-36
98 Blakeborough A, Ward J, Wilson D, et al. Hepatic lesion detection at MR imaging: a comparative study with four sequences. Radiology, 1997, 203(3): 759-765.
99 Weissleder R, Mahmood U. Molecular imaging. Radiology, 2001, 219(2): 316-333
100 Tjuvajev J G, Chen SH, Joshi A, et al. Imaging adenoviral mediated herpes virus thymidine kinase gene transfer and expression in vivo. Cancer Res, 1999, 59(20): 5186-5193
101 Encohs WS, Pet herick P, Bogdanova A, et al. Paramagnetic metal scavenging by melanin: MR imaging. Radiology, 1997, 204(2): 417-423
102 Weissleder R, Simonova M, Bogdanova A, et al. MR imaging and scintigrap hy of gene expression through melanin induction. Radiology, 1997, 204(2): 425-429
103 Ichikawa T, Hogemann D, Saeki Y. MRI of t ransgene expression: correlation to therapeutic gene expression. Neoplasia, 2002, 4(6): 523-530
104 Hogemann D, Ntziachristos V, Josephson L, et al. High throughput magnetic resonance imaging for evaluating targeted nanoparticle probes. Bioconjug Chem, 2002, 13(1): 116-121
105 Hogemann-Savellano D, Bos E, Blondet C, et al. The transferrin receptor: a potential molecular imaging marker for human cancer. Neoplasia, 2003, 5(6): 495-506
106 张卓然．实用细胞培养技术[M]．北京：人民卫生出版社，19991
107 Bulte JW, Duncan ID, Frank JA. In vivo magnetic resonance tracking of magnetically labeled ceils after transplantation. Journal of Cerebral Blood Flow & Metabolism. 2002, 22: 899-907
108 Josephson L, Tung CH, Morre A, et al. High-efficiency intracellular magnetic labeling with movel superparamagmetic-tat pettide conjugates. Bioconj Chem, 1999, 10: 186-191
109 Dodd CH, Hsu HC, Chu WJ, et al. Normal T-cell response and in vivo magnetic resonance imaging of T cells loaded with HIV transactivator-peptide-derived superparamagnetic nanoparticles. Immunol Methods, 2001, 256: 89-105.
110 扈盛，李世普，闫玉华，等．无机纳米粒子进入后肝癌细胞超微结构的变化．武汉理工大学学报，2005，27(11)：21-23