摘要
生物相容和生物可降解磁性纳米球是由具有生物相容性和生物可降解性的物质包覆纳米磁性材料制备而成的,它不但具有高分子磁性纳米球的多功能基团性和强磁性,而且由于其壳层材料是天然绿色的材料,因此它比普通高分子磁性纳米球有更广阔的应用前景,尤其是在医学领域。本文基于磁靶向纳米给药系统,采用两种新颖的、绿色的方法制备出了两种生物相容和生物可降解磁性纳米球,并对制备工艺进行优化,得到各自的较优制备方案,同时还详尽阐述了制备机理,具体内容如下:
(1)以油酸(OA)与十一烯酸(UA)共同修饰的Fe304为磁核,环已烷为中介,牛血清蛋白(BSA)为壳层组分,创新地用超声化学法制备了牛血清蛋白包覆Fe304(Fe3O4/BSA)磁性纳米球。着重探讨了BSA用量、磁流体浓度和用量、超声时间、超声功率、预乳化和水冷等因素对实验的影响,并获得了较优制备方案。在此方案下制备的样品通过FT-IR、TEM、SEM、TG以及VSM等分析手段进行了表征,结果表明纳米球壳层明显,粒径小(约100nm),成球性和分散性好,官能基团多,磁感应性较强(饱和磁化强度为38.5 emu/g),且具有超顺磁性,纳米球中Fe304的含量为57.6%。本文对超声化学法制备Fe3O4/BSA磁性纳米球的形成机理进行了详细分析和阐述,并通过对比实验进行了验证。
(2)对低温水热法制备含碳多聚糖微球和含碳多聚糖包覆Fe3O4(Fe3O4@CP)磁性纳米球的方法进行了改进,以微量NaOH为催化剂,产率和包覆效果都大幅提高。对此本文同样也获得了较优制备方案,在此方案下制备的Fe3O4@CP磁性纳米球,通过VSM分析可知,其饱和磁化强度高达60.629 emu/g;通过红外分析可知,纳米球壳层为含碳多聚糖,具有丰富的羟基和羰基;从热失重曲线可看出,纳米球的Fe304含量很高,达到81.7%;TEM图显示,纳米球的壳层明显,粒径在300nm以下。本文还证明了Fe304纳米粒子粒径较小时(小于20nm),用此法不能制备出以单个Fe304粒子为核的复合球体,而是形成无定型团聚体,并分析了原因。本文对含碳多聚糖微球以及Fe3O4@CP磁性纳米球的形成机理进行了解析,并提出了独到的见解。
Biocompatible and biodegradable magnetic nanospheres are synthesized by coating magnetic materials with biocompatible and biodegradable materials. They not only have surface functionality and high magnetic responsiveness as the polymer nanospheres, but also have shells made from natural and green materials, so their potential applications are much greater than normal polymer nanospheres, especially in medicine field. Magnetic nanospheres synthesized in this paper are based on magnetic targeted nanoparticle drug delivery system. Two kinds of biocompatible and biodegradable magnetic nanospheres were synthesized by two different novel methods in this paper. Their optimal synthesis conditions were got and their formation mechanisms were expounded. The details are described as below:
(1)Fe3O4 modified by OA and UA as the cores, bovine serum albumin (BSA) as the shells and cyclohexane as the agency, Fe3O4/BSA magnetic nanospheres were synthesized by sonochemical method. The effects of various factors in the experiments were studied, including BSA dosage, ferrofluid's concentration and dosage, ultrasonic time, ultrasonic power, pre-emulsification and water cooling. At last, optimal conditions were got. Coating effect and structure of the nanospheres synthesized under the optimal conditions were tested and characterized by FT-IR, TEM, SEM, TG and VSM. The results show that the nanospheres have clear core-shell structure, small size (approximately 100 nm), good sphericity and dispersibility, abundant functional groups, large saturated magnetization (38.5 emu/g) and superparamagnetism, high Fe3O4 content (57.6%).The formation mechanism of Fe3O4/BSA magnetic nanospheres synthesized by sonochemical method was analysed and expounded in detail, and it was proved through contrast experiments.
(2) Low-temperature hydrothermal method was modified by us. With this method, carbonaceous polysaccharide microspheres and carbonaceous polysaccharide coated Fe3O4 (Fe3O4@CP) magnetic nanospheres were synthesized. It was discovered that the yield and coating effect will be improved with the help of little NaOH. The optimal conditions were also found through a lot of experiments. Various properties of the Fe3O4@CP magnetic nanospheres synthesized under the optimal conditions were tested and characterized. They showed high saturated magnetization (60.629 emu/g) in the VSM test, high Fe3O4 content (81.7%) in the TG test. The TEM images show that their particle size is less than 300 nm. It was demonstrated that Fe3O4@CP magnetic nanospheres can not be synthesized with hydrothermal method but unformed agglomerates when small size Fe3O4 particles (less than 20 nm) were used as the cores, and the reasons were analysed. The formation mechanisms of carbonaceous polysaccharide microspheres and Fe3O4@CP magnetic nanospheres were inferred, and unique viewpoints were put forward in the paper.
引文
[1]Moghimi S M, Rajabi-Siahboomi A R. Recent advance in celllar, sub-cellular and molecular[J].Advance of Drug Delivery Review,2000,41:129~132
[2]Tartaj P, Morales M P, et al.Advances in magneyic nanoparticles for biotechnology applications[J].Journal of magnetism and Magnetic Materials,2005,290-291:28~34
[3]Tanaka T, Matsunaga T. Fully automated chemiluminescence immunoassay of insulin using antibody-protein a-bacterial magnetic particle complexes[J].Analytical Chemistry,2000,72:3518~3522
[4]宋海峰.磁性药物靶向治疗肿瘤[J].国外医学肿瘤学分册,2002,(3):183~186
[5]胡书春,周祚万.磁性高分子微球研究进展[J].材料科学与工程学报,2003,21,(4):616~619
[6]Hafeli U O. Magnetically modulated therapeutic system[J].International Journal of Pharmaceutics,2004,277:19~24
[7]Yang J.,Park S.B.,Yoon Ho-Geun, et al.Preparation of poly ε-caprolactone nanoparticles containing magnetite for magnetic drug carrier[J].International Journal of Pharmaceutics,2006,324:185~190
[8]Sachin Shaw, Murthy P.V.S.N.,Pradhan S.C..Effect of non-Newtonian characteristics of blood on magnetic targeting in the impermeable micro-vessel[J].Journal of Magnetism and Magnetic Materials,2010,322:1037~1043
[9]朱瀛,陆伟根.磁性微球和磁性纳米粒的研究进展[J].中国医药工业杂志,2005,36(5):581~584
[10]Kawashima Yoshiaki.Nanoparticulate systems for improved drug delivery[J].Advanced Drug Delivery Reviews,2001,47(1):1-2
[11]Marianne B, Gerhard S, Edmund P, et al.Cystic adventitial degeneration of the popliteal artery-the diagnostic value of duplex sonography[J].European Journal of Radiology, 2001,38(3):209~212
[12]马小军.单分散聚苯乙烯微球的制备及其磁化过程初探.硕士学位论文[D].中国科学院大连化学物理研究所,2004
[13]Hafeli U O.,Sweeny S M, Beresford B A, et al.Effective targeting of magnetic radioactive 90Y-microspheres to tumor cells by an externally applied magnetic field[J]. Nuclear Medicine and Biology,1995,22:147~155
[14]Russ Knapp F. F.,Mirzadeh S.,Beets A. L.,et al.Reactor-produced radioisotopes from ORNL for bone pain palliation[J].Applied Radiation and Isotopes,1998,4(49): 309~315
[15]Junfeng Y, Hafeli U O.,Dong Y, et al.Radiolabeling of magnetic targeted carriers with several therapeutic and imaging radioisotopes[J].European Cells & Materials Journal, 2002,3(2 Suppl):16~18
[16]李贵平,汪勇先,张春富,等.188Re标记免疫靶向磁性微粒及其生物学分布[J].中华核医学杂志,2006,26(4):231~235
[17]李贵平,汪勇先,张春富,等.抗癌胚抗原单抗免疫磁性纳米微粒的制备及其与188Re的放射性标记[J].放射性免疫学杂志,2005,18(6):410~412
[18]Zhang Chunfu, Cao Jinquan, Yin Duanzhi, et al.Preparation and radiolabeling of human serum albumin (HSA)-coated magnetite nanoparticles for magnetically targeted therapy[J].Applied Radiation and Isotopes,2004,61:1255~1259
[19]Bee A, Massart R, et al. Syntheises of very fine maghemite partieles[J].Journal of Magnetism and Magnetic Materials,1995,149:6-9
[20]李乾锋,刘颖,赵修臣.沉淀氧化法制备Fe304磁性粒子的研究[J].材料与表面处理技术,2006,6:63~65
[21]Xie Jin, Chen Kai, Lee Ha-Young, et al.Ultrasmall c(RGDyK)-coated Fe3O4 nanoparticles and their specific targeting to integrin αvβ3-rich tumor cells[J].Journal of the American Chemical Society,2008,130:7542~7543
[22]Fauconnier N.,Pons J. N.,Roger J.,et al. Thiolation of maghemite nanoparticles by dimercaptosuccinic acid[J].Journal of Colloid and Interface Science,1997,194: 427~433
[23]Jun Young-wook, Huh Yong-Min, Choi Jin-sil, et al.Nanoscale size effect of magnetic nanocrystals and their utilization for cancer diagnosis via magnetic resonance imaging[J].Journal of the American Chemical Society,2005,127:5732~5733
[24]Sun Shouheng, Zeng Hao, David B.Robinson, et al.Monodisperse MFe2O4 (M=Fe, Co, Mn) nanoparticles[J].Journal of the American Chemical Society,2004,126: 273~279
[25]Jia B P, Gao L. Fabrication of "tadpole"-like magnetite/multiwalled carbon nanotube heterojunctions and their self-assembly under external magnetic field[J].J. Phys. Chem. B,2007,111:5337~5343
[26]王煦漫,张彩宁,古宏晨.包覆油酸的Fe304纳米粒子的制备[J].精细化工,2007,24(8):733~736
[27]施卫贤,杨俊,王亭杰,等.磁性Fe304微粒表面有机改性[J].物理化学学报,2001, 17(6):507~510
[28]刘峥.Fe304微粒表面有机改性及其应用研究[J].化工技术与开发,2004,33(3):12~15
[29]Xu Lixing, Kim Min-Jung, Kim Ki-Do, et al.Surface modified Fe3O4 nanoparticles as a protein delivery vehicle[J].Colloids and Surfaces A:Physicochemical and Engineering Aspects,2009,350:8-12
[30]安丽娟,李兆强,俆娓,等.超顺磁性高分子微球的制备与表征[J].高等学校化学学报,2005,26(2):366~369
[31]蒋新宇,周春山,张俊山,等.导向药物用纳米Fe304磁性粒子的制备及表征[J].中南工业大学学报,2003,34(5):516~520
[32]Huh Yong-Min, Jun Young-wook, Song Ho-Taek, et al.In vivo magnetic resonance detection of cancer by using multifunctional magnetic nanocrystals[J].Journal of the American Chemical Society,2005,127:12387-12391
[33]Hila Cohen, Aharon Gedanken, Zhong Ziyi.One-step synthesis and characterization of ultrastable and amorphous FesO4 colloids capped with cysteine molecules[J].The Journal of Physical Chemistry C,2008,112:15429~15438
[34]Shieh Dar-Bin, Cheng Fong-Yu, Su Chia-Hao, et al. Aqueous dispersions of magnetite nanoparticles with NH3+ surfaces for magnetic manipulations of biomolecules and MRI contrast agents[J].Biomaterials,2005,26:7183~7191
[35]Fabio Sonvico, Stephane Mornet, Sebastien Vasseur, et al. Folate-conjugated iron oxide nanoparticles for solid tumor targeting as potential specific magnetic hyperthermia mediators:synthesis, physicochemical characterization, and in vitro experiments[J]. Bioconjugate Chemistry,2005,16:1181~1188
[36]汤健星.MRI造影剂介绍[J].医用放射技术杂志,2003,9:32
[37]刘学兵,王志刚,许川山,等.纳米级靶向超声造影剂的制备及对兔VX2肝肿瘤的显影试验[J].中国超声医学杂志,2009,1(25):5-8
[38]谢钢,张秋禹,李铁虎.磁性高分子微球[J].高分子通报,2001,(6):38~39
[39]Keziban Can, Mustafa Ozmen, Mustafa Ersoz. Immobilization of albumin on aminosilane modified superparamagnetic magnetite nanoparticles and its characterization[J].Colloids and Surfaces B:Biointerfaces,2009,71:154~159
[40]Cui Sheng, Shen Xiaodong, Lin Benlan. Surface organic modification of Fe3O4 nanoparticles by silane-coupling agents[J].Rare Metals,2006,25:426~430
[41]Masaki Uchida, Michelle L. Flenniken, Mark Allen, et al.Targeting of cancer cells with ferrimagnetic ferritin cage nanoparticles[J].J. Am. Chem. Soc.,2006,128: 16626~16633
[42]Rosenberg M.,Lee S.J..Water-insoluble, whey protein-based microspheres prepared by an all-aqueous process[J].Journal of Food Science,2004,1(64):50~58
[43]Lucie Beaulieu, Laurent Savoie, Paul Paquin, et al.Elaboration and characterization of whey protein beads by an emulsification/cold gelation process:application for the protection of retinol[J].Biomacromolecules,2002,3:239~248
[44]Suslick Kenneth S.,Grinstaff Mark W.. Protein microencapsulation of nonaqueous liquids[J].J.Am.Chem. Soc.,1990,112:7807~7809
[45]Grinstaff Mark W.,Suslick Kenneth S..Air-filled proteinaceous microbubbles: synthesis of an echo-contrast agent[J].Proc. Natl.Acad. Sci.USA,1991,(88): 7708~7710
[46]Avivi(Levi) S.,Nitzan Y.,Dror R.,et al. An easy sonochemical route for the encapsulation of tetracycline in bovine serum albumin microspheres[J].J. Am.Chem. Soc.,2003,125:15712~15713
[47]Farah Jean-Jacques Toublan, Stephen Boppart, and Suslick Kenneth S..Tumor targeting by surface-modified protein microspheres[J].J.Am.Chem. Soc.,2006,128:3472~3473
[48]Joe Z. Sostaric, Ramasamy P. Pandian, Anna Bratasz, et al.Encapsulation of a highly sensitive EPR active oxygen probe into sonochemically prepared microspheres[J].J. Phys. Chem. B,2007,111:3298~3303
[49]马建华,陈道达,田源,等.阿霉素磁性蛋白微球的改良法制备[J].中华实验外科杂志,2002,3(19):272~273
[50]邱广亮,李咏兰,邱广明,等.肺靶向顺铂磁性白蛋白微球的制备及性质[J].中国医药工业杂质,2001,32(12):544~546
[51]彦秋平,李富荣,汤顺清,等.免疫磁性海藻酸钠载药纳米微球的制备与评价[J].中国生物医学工程学报,2006,3(25):305~309
[52]Gorka Orive, Maria De Castro, Kong Hyun-Joon, et al.Bioactive cell-hydrogel microcapsules for cell-based drug delivery[J].Journal of Controlled Release,2009,135: 203~210
[53]刘善奎,高申,钟延强,等.DNA疫苗海藻酸钠微球的制备及体外释药[J].第二军医大学学报,2004,25(1):58~60
[54]Ainhoa Murua, Gorka Orive, Rosa M. Hernandez, et al. Xenogeneic transplantation of erythropoietin-secreting cells immobilized in microcapsules using transient immunosuppression[J].Journal of Controlled Release,2009, xxx:xxx-xxx
[55]Giovanni luca, Mario Calvitti,Claudio Nastruzzi, et al. Encapsulation, in vitro characterization, and in vivo biocompatibility of sertoli cells in alginate-based microcapsules[J].Tissue Enginering,2007,3(13):641~648
[56]Yoon Yeo, Kinam Park. A new microencapsulation method using an ultrasonic atomizer based on interfacial solvent exchange[J].Journal of Controlled Release,2004, 100:379~388
[57]Park Jae Hyung, Ye Mingli, Yeo Yoon, et al.Reservoir-type microcapsules prepared by the solvent exchange method:effect of formulation parameters on microencapsulation of lysozyme[J].Molecular Pharmaceutics,2005,2(3):135~143
[58]Kang Jichao, Schwendeman Steven P.Pore closing and opening in biodegradable polymers and their effect on the controlled release of proteins[J].Molecular Pharmaceutics,2006,1(4):104~118
[59]Saatchi Katayoun and Hafeli Urs O..Radiolabeling of biodegradable polymeric microspheres with [99mTc(CO)3]+ and in vivo biodistribution evaluation using microSPECT/CT imaging[J].Bioconjugate Chem.,2009,20:1209~1217
[60]Zhao Hong, KatayounSaatchi, Hafeli Urs O.,et al.Preparation of biodegradable magnetic microspheres with poly(lactic acid)-coated magnetite[J].Journal of Magnetism and Magnetic Materials,2009,321:1356~1363
[61]Hafeli Urs, Pauer Gayle, Failing Sarah, et al. Radiolabeling of magnetic particles with rhenium-188 for cancer therapy[J].Journal of Magnetism and Magnetic Materials,2001, 225:73~78
[62]Li Yan, Xu Xiuqing, Qi Dawei,et al.Novel Fe3O4@TiO2 core-shell microspheres for selective enrichment of phosphopeptides in phosphoproteome analysis[J].Journal of Proteome Research,2008,7:2526~2538
[63]邹东娜,张典瑞,杨海峰.白蛋白作为靶向给药载体的研究[J].食品与药品,2007,9(1):54~56
[64]韩萍芳,陈腊梅,姚成,等.超声对蛋白质盐析作用的研究[J].应用声学,2006,25(2):116~120
[65]王凯,马光辉.白蛋白微球的制备、改性和应用的研究进展[J].国外医学药学分册,2003,30(6):366~371
[66]陈锋,李春忠.改良热固法制备白蛋白微球的工艺研究[J].化学试剂,2005,27(6):365~367
[67]孟祥,李保国,齐正,吴亚卿.茶多酚磁性白蛋白微球的制备[J].食品科技,2007,1:47~49
[68]崔升,沈晓冬,林本兰,等.纳米磁性阿霉素人血白蛋白微球的制备与表征[J].中 国生物医学工程学报,2006,25(1):114~116
[69]Bumgarner Gary W.,Rangaiah Shashidharamurthy, Shanmugam Nagarajan, et al. Surface engineering of microparticles by novel protein transfer for targeted antigen/drug delivery[J].Journal of Controlled Release,2009, xxx:xxx-xxx
[70]Coombes A.G.A.,Breeze V.,Wu Lin,et al.Lactic acid-stabilised albumin for microsphere formulation and biomedical coatings[J].Biomaterials,2001,22:1-8
[71]Wang Jinyi, Wang Xueqin, Ren Li, et al.Conjugation of biomolecules with magnetic protein microspheres for the assay of early biomarkers associated with acute myocardial infarction[J].Anal.Chem.,XXXX,xxx,000-000
[72]Suslick Kenneth S..Applications of ultrasound to materials chemistry[J].Annu. Rev. Mater. Sci.,1999,29:295~326
[73]Morel Anne-Laure, Nikitenko Sergei I.,Gionnet Karine, et al.Sonochemical approach to the synthesis of Fe3O4@SiO2 core-shell nanoparticles with tunable properties[J].ACS Nano,2008,2(5):847~856
[74]Bunker Christopher E.,Novak Kyle C.,Guliants Elena A.,et al.Formation of protein-metal oxide nanostructures by the sonochemical method:observation of nanofibers and nanoneedles[J].Langmuir,2007,23:10342~10347
[75]Deng Shenglou, Umesh Gangadharmath, and Cheng-Wei Tom Chang. Sonochemistry: a powerful way of enhancing the efficiency of carbohydrate synthesis[J].J. Org. Chem., 2006,71:5179~5185
[76]Suslick Kenneth S.,Fang Mingming, and Taeghwan Hyeon. Sonochemical synthesis of iron colloids[J].J. Am.Chem. Soc.,1996,118:11960~11961
[77]Liu Ke Jian, Grinstaff Mark W.,Jiang Jinjie, et al.In vivo measurement of oxygen concentration using sonochemically synthesized microspheres[J].Biophysical Journal, 1994,67:896~901
[78]Olga Grinberg, Michal Hayun, Benjamin Sredni, et al.Characterization and activity of sonochemically-prepared BSA microspheres containing Taxol-An anticancer drug[J]. Ultrasonics Sonochemistry,2007,14:661~666
[79]Olga Grinberg, Aharon Gedanken, Chitta Ranjan Patra, et al.Sonochemically prepared BSA microspheres containing Gemcitabine, and their potential application in renal cancer therapeutics[J].Acta Biomaterialia,2009,5:3031~3037
[80]Zhou Meifang, Thomas Seak Hou Leong, Sonia Melino, et al. Sonochemical synthesis of liquid-encapsulated lysozyme microspheres[J].Ultrasonics Sonochemistry,2010,17: 333~337
[81]Avivi(Levi) S.,Gedanken A.. The preparation of avidin microspheres using the sonochemical method and the interaction of the microspheres with biotin[J].Ultrasonics Sonochemistry,2005,12:405~409
[82]王镜岩,朱圣庚,徐长法.生物化学[M].第三版(上册).北京:高等教育出版社,2002.97~99
[83]Hu Bo, Wang Kan, Wu Liheng, et al. Engineering carbon materials from the hydrothermal carbonization process of biomass[J].Adv. Mater.,2010,22:813~828
[84]Sun Xiaoming and Li Yadong. Colloidal carbon spheres and their core/shell structures with noble-metal nanoparticles[J].Angew. Chem. Int. Ed.,2004,43:597-601
[85]Wang Qing, H Li ong, Chen Liquan, et al.Monodispersed hard carbon spherules with uniform nanopores[J].Carbon,2001,39:2211~2214
[86]孙晓明.低维功能纳米材料的液相合成、表征与性能研究[D].清华大学博士学位论文,2005
[87]Emory S R, Haskins W E, Nie S M. Direct observation of size-dependent optical enhancement in single metal nanoparticles[J].J. Am. Chem. Soc.,1998,120: 8009~8010
[88]Chen Hemei, Deng Chunhui, Li Yan, et al. A facile synthesis approach to C8-functionalized magnetic carbonaceous polysaccharide microspheres for the highly efficient and rapid enrichment of peptides and direct MALDI-TOF-MS analysis[J].Adv. Mater.,2009,21:2200~2205
[89]Yao Guoping, Qi Dawei,Deng Chunhui, et al.Functionalized magnetic carbonaceous microspheres for trypsin immobilization and the application to fast proteolysis[J]. Journal of Chromatography A,2008,1215:82~91
[90]王镜岩,朱圣庚,徐长法.生物化学[M].第三版(上册).北京:高等教育出版社,2002.36
[91]Gerard C.A.Luijkx, Fred van Rantwijk, Herman van Bekkum, et al.The role of deoxyhexonic acids in the hydrothermal decarboxylation of carbohydrates [J]. Carbohydrate Research,1995,272:191~202
[92]李同起,王成扬.碳质中间相形成机理研究[J].新型炭材料,2005,20(3):278~285
[93]许凤华,尹端沚,汪勇先.188Re标记生物分子研究进展[J].核技术,2008,12(31):931~937
[94]马玉飞,于俊峰,周伟,等.铼羰基化合物标记含RGD的多肽[J].核化学与放射化学,2009,1(31):23~27
[95]唐林,于俊峰,夏姣云,等.羰基铼[188Re]标记含RGD多肽[J].同位素,2006, 1(19):17~21
[96]夏姣云,汪勇先,于俊峰,等.三羰基铼[188Re]的放射化学合成[J].核化学与放射化学,2005,2(27):87~90
[97]曹金全,汪勇先,于俊峰,等.磁性纳米微粒的188Re标记[J].同位素,2004,2(17):84~89
[98]Cao Jinquan, Wang Yongxian, Yu Junfeng, et al.Preparation and radiolabeling of surface-modified magnetic nanoparticles with rhenium-188 for magnetic targeted radiotherapy[J].Journal of Magnetism and Magnetic Materials,2004,277:165~174
[99]Benjamin B.Yellen, Zachary G. Forbes, Derek S.Halverson, et al. Targeted drug delivery to magnetic implants for therapeutic applications[J].Journal of Magnetism and Magnetic Materials,2005,293:647~654