PLGA/O-CMC抗癌药物及基因复合功能纳米载体系统的构建及评价
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摘要
本文以聚乳酸-乙醇酸共聚物（PLGA）和自行制备的 O-羧甲基壳聚糖（O-CMC）
为原料，分别以 5-氟尿嘧啶为抗癌药物模型，以反义 EGFR 为基因药物模型，采
用自身设计的改良复乳法制备了载药和载基因纳米微粒；并在此基础上构建与评
价了同载抗癌药物与基因的复合功能纳米药物载体系统。
各种纳米微粒表征结果显示：（1）以 O-CMC 和 PLGA 为原料制得的空载纳米
粒子平均粒径为 222.9nm，粒径分布指数为 0.109，ζ电位为 73.46eV，表面氮
元素含量为 10.3%，具有明显的核壳结构和良好的表面亲水性；（2）以 5-氟尿嘧
啶（5-FU）为抗癌药物模型制得的载药纳米微粒平均粒径为 98.5nm，粒径分布
指数为 0.192，粒子表面ξ电位为 61.48eV，表面 N 元素含量为 12.2%，对 5-FU
的包载率高达 18.9%；（3）以反义 EGFR 为基因药物模型制得的表面吸附加内部
包载基因的纳米微粒平均粒径为 375.3nm，粒径分布指数为 0.263，ζ电位为
-10.67eV。
本文用 SEM 动态监测各种纳米粒子降解过程中表面形貌的变化，并连续追踪
各种纳米粒子降解过程中的质量损失和降解介质的 pH 变化。载 5-FU 纳米粒子
在 PBS 中的释药行为研究表明：（1）前 12 小时的释药动力学符合 Huguchi 方程，
具有一级释放特性；（2）在 20 天内的释药动力学符合零级释放特性。作者创新
构建的表面吸附加内部包载基因的纳米微粒释药动力学方程也具有零级释放动
力学特性。
体外细胞存活率实验和体内动物实验均证实了 PLGA/O-CMC 纳米粒子有较好
的生物相容性；MTT 和细胞凋亡实验结果表明载药纳米粒子和载基因纳米微粒
对 TJ905 脑胶质瘤细胞增殖有明显的抑制作用；免疫组化实验进一步证实包载基
因的纳米微粒能有效地抑制胶质瘤细胞中 EGFR 的表达，从而抑制 TJ905 人脑胶
质瘤细胞的增殖。最后用荧光相差显微镜和共聚焦显微镜动态监测了纳米粒子的
细胞转染过程，发现纳米粒子可在不同时间内进入细胞浆和细胞核。
本文构建的表面吸附基因且内部同时包载 5-FU 与基因的复合功能纳米微粒
对基因的包载量显著提高，并对肿瘤细胞有较高的抑制率，表明复合功能纳米微
粒中各组分对肿瘤细胞增殖的抑制具有明显的协同作用。
The traditional double emulsion (water-in-oil-in-water) method to prepare
hydrophilic drug-loaded nanoparticles (NPs) was modified and improved to get
smaller NPs and high encapsulation efficiency of drug; Poly (D, L-lactide-co-glycolic
acid) (PLGA) and o-carboxymethyl chitosan (O-CMC) were chosen to
microencapsulate 5-fluorouracil (5-FU) and antisense-EGFR cDNA by improved
W/O/W method due to their attractive degradation and physicochemical properties;
So 5-FU and antisense-EGFR-loaded PLGA/O-CMC NPs were created to realize the
combination of anticancer drug and gene in one kind of biodegradable nanocarrier.
The results of characterizing all kinds of nanoparticles showed that: (1) mean
size of O-CMC-coated PLGA NPs without drug is 222.9nm, polydispersity is 0.109,
Zeta potenial is 73.46eV, N percentage on the surface of NPs is 10.3%, hydrophilicity
of NPs is very good; （2）mean size of 5-FU-coated PLGA/O-CMC NPs is 98.5nm,
polydispersity is 0.192, Zeta potenial is 61.48eV, N percentage on the surface of NPs
is 12.2%,5-FU-loading level is 18.9%; (3) mean size of antisense-EGFR-encapsulated
PLGA/O-CMC NPs while absorbing antisense-EGFR is 375.3nm, polydispersity is
0263, Zeta potenial is –10.67eV.
SEM was used to study the morphological change of NPs during degradation, and
weight loss of NPs and pH change of degradation medium were traced. Researches of
5-FU release behavior from NPs showed that (1) release kinetics of 5-FU from NPs in
early 12 hours was coincidence with Huguchi release; （2）release kinetics in 20 days
was coincidence with Zero-level release. Release kinetics of
antisense-EGFR-encapsulated PLGA/O-CMC NPs while absorbing antisense-EGFR
was accordance with Zero-level release.
Cell viability in vitro as well as in vivo demonstrated that biocompatibility of
PLGA/O-CMC NPs was good. Results of MTT and cell apoptosis illustrated that both
5-FU-encapsulated NPs and antisense-EGFR-loaded NPs had high cytotoxicity on
brain tumor cells TJ905, and immunohistochemical staining approved that
antisense-EGFR-loaded NPs had inhibitory effect on the expression of EGFR of
human gliomas cells. Fluorescence microscope and confocal microscope were used to
trace the procedures of cell transfection induced by NPs. It was found that there were
some NPs were in nucleus after 24h.
Both 5-FU and antisense-EGFR-encapsulated NPs while absorbing EGFR on the
surface of NPs designed by us creatively had high antisense-EGFR loading level and
high cytotoxicity on gliomas cells (73.6%), which showed that 5-FU and
antisense-EGFR had cooperation effect on the inhibitory effect of gliomas cells.
为原料，分别以 5-氟尿嘧啶为抗癌药物模型，以反义 EGFR 为基因药物模型，采
用自身设计的改良复乳法制备了载药和载基因纳米微粒；并在此基础上构建与评
价了同载抗癌药物与基因的复合功能纳米药物载体系统。
各种纳米微粒表征结果显示：（1）以 O-CMC 和 PLGA 为原料制得的空载纳米
粒子平均粒径为 222.9nm，粒径分布指数为 0.109，ζ电位为 73.46eV，表面氮
元素含量为 10.3%，具有明显的核壳结构和良好的表面亲水性；（2）以 5-氟尿嘧
啶（5-FU）为抗癌药物模型制得的载药纳米微粒平均粒径为 98.5nm，粒径分布
指数为 0.192，粒子表面ξ电位为 61.48eV，表面 N 元素含量为 12.2%，对 5-FU
的包载率高达 18.9%；（3）以反义 EGFR 为基因药物模型制得的表面吸附加内部
包载基因的纳米微粒平均粒径为 375.3nm，粒径分布指数为 0.263，ζ电位为
-10.67eV。
本文用 SEM 动态监测各种纳米粒子降解过程中表面形貌的变化，并连续追踪
各种纳米粒子降解过程中的质量损失和降解介质的 pH 变化。载 5-FU 纳米粒子
在 PBS 中的释药行为研究表明：（1）前 12 小时的释药动力学符合 Huguchi 方程，
具有一级释放特性；（2）在 20 天内的释药动力学符合零级释放特性。作者创新
构建的表面吸附加内部包载基因的纳米微粒释药动力学方程也具有零级释放动
力学特性。
体外细胞存活率实验和体内动物实验均证实了 PLGA/O-CMC 纳米粒子有较好
的生物相容性；MTT 和细胞凋亡实验结果表明载药纳米粒子和载基因纳米微粒
对 TJ905 脑胶质瘤细胞增殖有明显的抑制作用；免疫组化实验进一步证实包载基
因的纳米微粒能有效地抑制胶质瘤细胞中 EGFR 的表达，从而抑制 TJ905 人脑胶
质瘤细胞的增殖。最后用荧光相差显微镜和共聚焦显微镜动态监测了纳米粒子的
细胞转染过程，发现纳米粒子可在不同时间内进入细胞浆和细胞核。
本文构建的表面吸附基因且内部同时包载 5-FU 与基因的复合功能纳米微粒
对基因的包载量显著提高，并对肿瘤细胞有较高的抑制率，表明复合功能纳米微
粒中各组分对肿瘤细胞增殖的抑制具有明显的协同作用。
The traditional double emulsion (water-in-oil-in-water) method to prepare
hydrophilic drug-loaded nanoparticles (NPs) was modified and improved to get
smaller NPs and high encapsulation efficiency of drug; Poly (D, L-lactide-co-glycolic
acid) (PLGA) and o-carboxymethyl chitosan (O-CMC) were chosen to
microencapsulate 5-fluorouracil (5-FU) and antisense-EGFR cDNA by improved
W/O/W method due to their attractive degradation and physicochemical properties;
So 5-FU and antisense-EGFR-loaded PLGA/O-CMC NPs were created to realize the
combination of anticancer drug and gene in one kind of biodegradable nanocarrier.
The results of characterizing all kinds of nanoparticles showed that: (1) mean
size of O-CMC-coated PLGA NPs without drug is 222.9nm, polydispersity is 0.109,
Zeta potenial is 73.46eV, N percentage on the surface of NPs is 10.3%, hydrophilicity
of NPs is very good; （2）mean size of 5-FU-coated PLGA/O-CMC NPs is 98.5nm,
polydispersity is 0.192, Zeta potenial is 61.48eV, N percentage on the surface of NPs
is 12.2%,5-FU-loading level is 18.9%; (3) mean size of antisense-EGFR-encapsulated
PLGA/O-CMC NPs while absorbing antisense-EGFR is 375.3nm, polydispersity is
0263, Zeta potenial is –10.67eV.
SEM was used to study the morphological change of NPs during degradation, and
weight loss of NPs and pH change of degradation medium were traced. Researches of
5-FU release behavior from NPs showed that (1) release kinetics of 5-FU from NPs in
early 12 hours was coincidence with Huguchi release; （2）release kinetics in 20 days
was coincidence with Zero-level release. Release kinetics of
antisense-EGFR-encapsulated PLGA/O-CMC NPs while absorbing antisense-EGFR
was accordance with Zero-level release.
Cell viability in vitro as well as in vivo demonstrated that biocompatibility of
PLGA/O-CMC NPs was good. Results of MTT and cell apoptosis illustrated that both
5-FU-encapsulated NPs and antisense-EGFR-loaded NPs had high cytotoxicity on
brain tumor cells TJ905, and immunohistochemical staining approved that
antisense-EGFR-loaded NPs had inhibitory effect on the expression of EGFR of
human gliomas cells. Fluorescence microscope and confocal microscope were used to
trace the procedures of cell transfection induced by NPs. It was found that there were
some NPs were in nucleus after 24h.
Both 5-FU and antisense-EGFR-encapsulated NPs while absorbing EGFR on the
surface of NPs designed by us creatively had high antisense-EGFR loading level and
high cytotoxicity on gliomas cells (73.6%), which showed that 5-FU and
antisense-EGFR had cooperation effect on the inhibitory effect of gliomas cells.
引文
参考文献
参考文献
[1] 王身国，蔡晴，吕泽，贝建中，Chinese J. Reparative and Reconstructive Surgery,
2001, 15(5): 280-285
[2] R.A. Muzzarelli, C. Jeunmux, and G. W. Gooday. Plenum Press, New York, 1986
[3] Muazzrelli R. A. A. Pergamon Press, 1977
[4] Handbook of Chemistry and Physis, 55th Edition, P.1974-1975
[5] Wintemantel E, Mayer J., Blum J., et al. Biomaterials, 1996, 17(2): 83
[6] Dunn RL, In Hollinger J. Boca Raton, CRC Press, Florida, 1995:1731
[7] Iznmikawa S., Yoshioka S., Aso Y, et al. J. Control. Release, 1991, 15:133
[8] Pitt CG，Langer R, Chasin M Eds，Marcel Dekker, New York, 1990: 71105
[9] Li SM, Vent M. G. Scott and D.Filead eds. Chapman & Hall, London, 1995:4387
[10] Langer R. ACC Chem Res, 2000, 33(2): 94
[11] Leong KW, Brott BC, Langer R, J Biomed Mater Res, 1985, 19: 941
[12] Langer R. Nature, 1998, 392: 5
[13] Domb AJ, Langer R, U. S. Patent 4, 757, 128 (Jul, 12, 1988)
[14] Chichering D, Jacob J, Mathiowitz E. Biotechnology and Bioengineering, 1996,
52: 96
[15] Domb AJ, Maniar M. J Polym Sci, Part A: Polym Chem, 1993, 31: 1275
[16] Domb AJ, Nudelman R. J Polym Sci, Part A: Polym Chem, 1995, 33: 717
[17] Domb AJ, Lsrael ZH, Elmalak O, et al. Pharm Res 1999, 16(5): 762
[18] Thomasin C, Corradin G, Men Y, et al. J Controlled Release, 1996, 41: 131-145
[19] Bleich J, Mueller BW. Wassmus W. Int J Pharm, 1993, 97(1-3): 111-117
[20] Hayashi Y, Yoshioka S, Aso Y, et al. Pharm Res, 1994, 11(2): 337-340
[21] Nihant N. Grandfils C. Jerome R, et al. J Controlled Release, 1995, 35(2-3):
117-125
[22] McGee JP, Davis SS, O’Hagan DT. J controlled Release, 1995, 34(2): 77-86
[23] Ruiz JM, Tissier B, Benoit JP. Int J Pharm, 1989, 49(1): 69-77
103
参考文献 参考文献
[24] Park TG, Lee HY, Nam YS. J Controlled Release, 1998, 55(2-3): 181-191
[25] Mehta RC, Thanoo BC, Deluca PP. J Controlled Release, 1996, 41(3): 249-257
[26] Crotts G, Sah HK, Park, TG. J Controlled Release, 1997, 47(1): 101-111
[27] Li XH, Deng XM, Yuan ML, et al. Int J Pharm, 1999, 178(2): 245-255
[28] Uchida T, Yoshida K, Ninomiya A, et al. Chem Pharm Bull, 1995, 43(9): 1569-1573
[29] Rojas J, Pinto-Alphandary H, Leo E, et al. Int J Pharm, 1999, 183(1): 67-71
[30] Rosa GD, Iommelli R, La Rotonda MI, et al. J controlled Release, 2000, 69(2):
283-295
[31] Joyanthi R, Thanoo BC, Metha RC, et al. J Controlled Release, 1996, 38: 235-244
[32] Yang YY, Chia HH, Chung TS, J Controlled Release, 2000, 69(1): 81-96
[33] Lourenco C, Teixeira M, Simoes S, et al. Int J Pharm, 1996, 138(12): 1
[34] Paul M, Fessi H, Laatiris A, et al. Int J Pharm, 1997, 159(15): 223
[35] Reich G. Drug Dev Ind Pharm, 1997, 23(12): 1191
[36] Landry FB, Bazile DV, Spenlehaver G, et al. STP Pharm Sci, 1996, 6(3): 195
[37] Landry FB, Bazile DV, Spenlehaver G, et al. J Controlled Release, 1997, 44(17):
227
[38] 王杰，张强，易翔，等，北京医科大学学报，2000，32（3）：235
[39] Leroux JC, Allamann E, Dedaeghere E, et al. Controlled Release, 1996, 39:339
[40] Soppimath KS, Aminabhavi TM, Kulkami AR, et al. J Controlled Release, 2001
70: 1-20
[41] P. Couvreur, L. Grislain , CRC Press, Boca Raton, FL, 1986: 27-93
[42] J. Kreuter, R.N, Alyautdin, D.A, Brain Res, 1995,674: 171-174
[43] G. Borchard, K.L. Audus, Int. J. Pharm, 1994:110 29-35
[44] Bennis. S., Robert, J., Nemati, F., Dubernet, C, et al. Cancer Res. 1992, 33,
A2846
[45] Couvreur ,C., Roblot-Treupel, L., Poupon, M.F., et al. Adv. Drug Delve. Rev.
1990, 5: 209-230
[46] Hu, Y.P., Toulme, N., Bennis, S. and Robert, J. Anticancer Drugs, 1994, 37-38
104
参考文献 参考文献
[47] Kreuter, J. Methods Enzymol., 1985, 112: 129-138
[48] Morin C, Barratt G, Fessi H, et al. Int J Immunopharmacol, 1994, 16: 451-456
[49] Godard, G., Boutorrine, A.S., Saison, B.E., et al. Eur. J. Biochem. 1995, 232:
404-410
[50] A Truong-Le, Vu L., Walsh, Scott M, et al. Arch. Biochem. Biophys. ,1999, 361(1):
47-56
[51] Godard G, Boutorine As, Saison BE, et al. Eur J Biochem. , 1995, 232(2), 404-410
[52] Kossovsky N, Gelman A, Hnatyszyn HJ, et al. Bioconjug Chem., 1995, 6: 507-511
[53] Stieneker, F., Kreuter, J., and Lower, J. AIDS, 1991, 5: 431-435
[54] Pinto-Alphandrary, H., Balland, O., Couvreur, P., et al. Pharm. Res. 1994,
11: 38-46
[55] Fattal, E., Rojas, J., Rolot, T.L.,et al. J. Microencapsulation 1991, 8: 29-36
[56] Lobenberg R, Kreuter J. AIDS Res Hum Retroviruses, 1996, 12(18): 1709-1715
[57] Stolnik S, Dunn S E, Illum L et al. Pharm. Res., 1994, 11: 1800-1808
[58] Gref R, Minamitake Y, Peracchia M T et al. Science, 1994, 263: 1600-1603
[59] Leroux J C, Cozens R, Roesel J L et al. J. Pharm. Sci., 1995, 84(12): 1387-1391
[60] Kim Y I, Fluckiger L, Hoffman M et al. Br. J. Pharmacol, 1997, 120(3): 399-404
[61] Damge C, Michel C, Aprahanian M, et al. Diabetes. , 1988, 37: 246
[62] Jani P, Halbert GW, Langridge J, et al. J Pharm Pharmacol ,1990, 42: 821
[63] Maincent, P., Le Verge, R., Couvreur, P. et al. J. Pharm. Sci.,1986, 75: 955-958
[64] Zimmer A, Mutschler E, Lambrecht G, et al. Pharm Res, 1994, 11(10): 1435-1442
[65] Kumaresh S, Tejraj M, J. Control. Release, 2001, 70:1-20
[66] Christine Allen a , Dusica, Colloids and Surfaces B: Biointerfaces ,1999, 16:
3-27
[67] 常津，刘海峰，许晓秋，中国生物医学工程学报，2002， 12
[68] 邵健，杨宇民，南通医学学报，1999
[69] Lingyun Chen, Yunmin Du, Xiaoqing Zeng, Carbohydrate Research，2003, 338：
333-340
[70] 陈洁，夏文水，日用化学工业，1999
105
参考文献 参考文献
[71] 罗彦凤，王远亮，高分子材料科学与工程，2003,19
[72] 何永言，夏淑贞，吴蒙等，高分子材料科学与工程，1993，2：24-28.
[73] 沈兴海 高宏成，化学通报，1995
[74] Sanjeeb K.Sahoo,Jayanth Panyam,et al,Journal of Controlled Release,
[75] L D Richard ,J O Hollinger eds.CRC Press, Inc.boca Raton.,1995: 17-31
[76] Gliding D K,Reed A M., Journal of Polymer,1979,20:1459
[77]VacantiJ.P.,MayerJ.E,Langer R.,et al.,Journal of annthorac
surg,1995,60:513-517
[78] Lexroux J.C.,Allamann E, Journal of Controlled Release,1996,39:339
[79] 赵锋，高永良，中国新药杂志，2002, 11
[80] 崔正刚，殷福珊，中国轻工业出版社，1999
[81] Lagourette B,Peyrelasse J,Boned C,et al., Nature,1979,281:60-62.
[82] 李晓，刘戈，袁惠根等，福州大学学报，2000，8
[83] 施利毅，张潘琪，高分子材料科学与工程，1999，7
[84] Gabriele Reich， Journal of Pharmaceutics and Biopharmaceutics， 1998，45：
165-171
[85] 赵锋，高永良，中国新药杂志，2002，11
[86] Schakenraad J.M.,Hardon M.J.,Feijen J.,et al., Journal of biomedical Materials
Research,1990,24:529-545.
[87] Bodmeier R.,Chen H., International Journal of Pharmartics,1989,51:1-8
[88] Vert M,Mauduit J,Li S.,biomaterials,1994,15:1209-1213
[89] Spenlehauer G,Vert M,Benoit J.P.,et,al.,biomaterials,1989,10:557-563
[90] Isobe M.,Yamazaki Y., Oida S-I., et al., Journal of biomedical Materials
Research, 1996,32:433-438
[91] Kamei S,Inoue Y., Advanced Polymer Science,1999,94:107-141
[92] James M.Anderson,Matthew S.Shive, Advanced Drug Delivery Reviews, 1997,
28:5-24
[93] 中华人民共和国药典二部（2000 版），北京，中国标准出版社，2000：472～474
[94] 沈正荣，朱家蕙，中国医药工业杂志，1996，28(7):306～308
[95] M.L.Hans,A.M.Lowman, 2002: 319-327
[96] Friederike von Burkersroda,Luise Schedl,Biomaterials,2002,23: 4221-4231
[97] 杨炳兴，成国祥，胶体与聚合物，2003
[98] L.B, Peppas,Inter. J. Pharm., 1995, 116:1～9
[99] Kerr J.R.,Harmon B.V.,Cancer,1994,73:2013
106
参考文献 参考文献
[100] Crystal R.G.,Science,1995,270～404
[101] Schafield J.P.,Caukey C.T.,Br Med Bull,1995,51(1):56～71
[102] Yamamoto T.,Kamata N.,Kawano H.,et al.,Cancer Res., 1986,44:414-416
[103] Michael F. Haller, Maryland， 2001
[104] 黄世文，卓仁禧，科学通报 48 卷 第五期,2003
[105] Cohen.L.S.,J.E.Rosenthal,D.W.Horner,American Journal of Cardiology
29:520-523
[106] Agrawal.C.M.and K.A.Athanasiou, J.of Biomed. Material. Research，1997，
38(2):105-114
[107] Eldridge.J.H.,C.J.Hammond, J. of Control. Release, 2:205-214
[108] Yamamoto T., Kamata N, Kawano H et al, Cancer Res., 1986,44:41-416
[109] Liberman T.A., Nusbaum H.R., Razon N., et al, Nature, 1985, 313: 14-147
[110] Luo D Saltzman M.,Synthetic,Natural Biotechnology,2000,18:33-37
[111] Nakanishi M.,Akuta T,Nagoshi E,et al.,European Journal Pharmartics
Science,2001.13:17-24
[112] Rolland A P.,Drug Delivery,1998,15:143-198
[113] Ferber B.,Science,2001,294:1638-1642
[114] Son K,Huang L., Proc Natl Acad Sci USA,1994,91:12669-12672
[115] Alexander IE, Russell DW, Miller AD., J Virol, 1994, 68:8282-8287
[116] Kuriyama S, Masui K, Sakamoto T, et al, Anticancer Res. 1998, 18:3399-3406.
[117] 陈刚 李世拥 于波，中华胃肠外科杂志，2002，1（5）：134－136
参考文献
[1] 王身国，蔡晴，吕泽，贝建中，Chinese J. Reparative and Reconstructive Surgery,
2001, 15(5): 280-285
[2] R.A. Muzzarelli, C. Jeunmux, and G. W. Gooday. Plenum Press, New York, 1986
[3] Muazzrelli R. A. A. Pergamon Press, 1977
[4] Handbook of Chemistry and Physis, 55th Edition, P.1974-1975
[5] Wintemantel E, Mayer J., Blum J., et al. Biomaterials, 1996, 17(2): 83
[6] Dunn RL, In Hollinger J. Boca Raton, CRC Press, Florida, 1995:1731
[7] Iznmikawa S., Yoshioka S., Aso Y, et al. J. Control. Release, 1991, 15:133
[8] Pitt CG，Langer R, Chasin M Eds，Marcel Dekker, New York, 1990: 71105
[9] Li SM, Vent M. G. Scott and D.Filead eds. Chapman & Hall, London, 1995:4387
[10] Langer R. ACC Chem Res, 2000, 33(2): 94
[11] Leong KW, Brott BC, Langer R, J Biomed Mater Res, 1985, 19: 941
[12] Langer R. Nature, 1998, 392: 5
[13] Domb AJ, Langer R, U. S. Patent 4, 757, 128 (Jul, 12, 1988)
[14] Chichering D, Jacob J, Mathiowitz E. Biotechnology and Bioengineering, 1996,
52: 96
[15] Domb AJ, Maniar M. J Polym Sci, Part A: Polym Chem, 1993, 31: 1275
[16] Domb AJ, Nudelman R. J Polym Sci, Part A: Polym Chem, 1995, 33: 717
[17] Domb AJ, Lsrael ZH, Elmalak O, et al. Pharm Res 1999, 16(5): 762
[18] Thomasin C, Corradin G, Men Y, et al. J Controlled Release, 1996, 41: 131-145
[19] Bleich J, Mueller BW. Wassmus W. Int J Pharm, 1993, 97(1-3): 111-117
[20] Hayashi Y, Yoshioka S, Aso Y, et al. Pharm Res, 1994, 11(2): 337-340
[21] Nihant N. Grandfils C. Jerome R, et al. J Controlled Release, 1995, 35(2-3):
117-125
[22] McGee JP, Davis SS, O’Hagan DT. J controlled Release, 1995, 34(2): 77-86
[23] Ruiz JM, Tissier B, Benoit JP. Int J Pharm, 1989, 49(1): 69-77
103
参考文献 参考文献
[24] Park TG, Lee HY, Nam YS. J Controlled Release, 1998, 55(2-3): 181-191
[25] Mehta RC, Thanoo BC, Deluca PP. J Controlled Release, 1996, 41(3): 249-257
[26] Crotts G, Sah HK, Park, TG. J Controlled Release, 1997, 47(1): 101-111
[27] Li XH, Deng XM, Yuan ML, et al. Int J Pharm, 1999, 178(2): 245-255
[28] Uchida T, Yoshida K, Ninomiya A, et al. Chem Pharm Bull, 1995, 43(9): 1569-1573
[29] Rojas J, Pinto-Alphandary H, Leo E, et al. Int J Pharm, 1999, 183(1): 67-71
[30] Rosa GD, Iommelli R, La Rotonda MI, et al. J controlled Release, 2000, 69(2):
283-295
[31] Joyanthi R, Thanoo BC, Metha RC, et al. J Controlled Release, 1996, 38: 235-244
[32] Yang YY, Chia HH, Chung TS, J Controlled Release, 2000, 69(1): 81-96
[33] Lourenco C, Teixeira M, Simoes S, et al. Int J Pharm, 1996, 138(12): 1
[34] Paul M, Fessi H, Laatiris A, et al. Int J Pharm, 1997, 159(15): 223
[35] Reich G. Drug Dev Ind Pharm, 1997, 23(12): 1191
[36] Landry FB, Bazile DV, Spenlehaver G, et al. STP Pharm Sci, 1996, 6(3): 195
[37] Landry FB, Bazile DV, Spenlehaver G, et al. J Controlled Release, 1997, 44(17):
227
[38] 王杰，张强，易翔，等，北京医科大学学报，2000，32（3）：235
[39] Leroux JC, Allamann E, Dedaeghere E, et al. Controlled Release, 1996, 39:339
[40] Soppimath KS, Aminabhavi TM, Kulkami AR, et al. J Controlled Release, 2001
70: 1-20
[41] P. Couvreur, L. Grislain , CRC Press, Boca Raton, FL, 1986: 27-93
[42] J. Kreuter, R.N, Alyautdin, D.A, Brain Res, 1995,674: 171-174
[43] G. Borchard, K.L. Audus, Int. J. Pharm, 1994:110 29-35
[44] Bennis. S., Robert, J., Nemati, F., Dubernet, C, et al. Cancer Res. 1992, 33,
A2846
[45] Couvreur ,C., Roblot-Treupel, L., Poupon, M.F., et al. Adv. Drug Delve. Rev.
1990, 5: 209-230
[46] Hu, Y.P., Toulme, N., Bennis, S. and Robert, J. Anticancer Drugs, 1994, 37-38
104
参考文献 参考文献
[47] Kreuter, J. Methods Enzymol., 1985, 112: 129-138
[48] Morin C, Barratt G, Fessi H, et al. Int J Immunopharmacol, 1994, 16: 451-456
[49] Godard, G., Boutorrine, A.S., Saison, B.E., et al. Eur. J. Biochem. 1995, 232:
404-410
[50] A Truong-Le, Vu L., Walsh, Scott M, et al. Arch. Biochem. Biophys. ,1999, 361(1):
47-56
[51] Godard G, Boutorine As, Saison BE, et al. Eur J Biochem. , 1995, 232(2), 404-410
[52] Kossovsky N, Gelman A, Hnatyszyn HJ, et al. Bioconjug Chem., 1995, 6: 507-511
[53] Stieneker, F., Kreuter, J., and Lower, J. AIDS, 1991, 5: 431-435
[54] Pinto-Alphandrary, H., Balland, O., Couvreur, P., et al. Pharm. Res. 1994,
11: 38-46
[55] Fattal, E., Rojas, J., Rolot, T.L.,et al. J. Microencapsulation 1991, 8: 29-36
[56] Lobenberg R, Kreuter J. AIDS Res Hum Retroviruses, 1996, 12(18): 1709-1715
[57] Stolnik S, Dunn S E, Illum L et al. Pharm. Res., 1994, 11: 1800-1808
[58] Gref R, Minamitake Y, Peracchia M T et al. Science, 1994, 263: 1600-1603
[59] Leroux J C, Cozens R, Roesel J L et al. J. Pharm. Sci., 1995, 84(12): 1387-1391
[60] Kim Y I, Fluckiger L, Hoffman M et al. Br. J. Pharmacol, 1997, 120(3): 399-404
[61] Damge C, Michel C, Aprahanian M, et al. Diabetes. , 1988, 37: 246
[62] Jani P, Halbert GW, Langridge J, et al. J Pharm Pharmacol ,1990, 42: 821
[63] Maincent, P., Le Verge, R., Couvreur, P. et al. J. Pharm. Sci.,1986, 75: 955-958
[64] Zimmer A, Mutschler E, Lambrecht G, et al. Pharm Res, 1994, 11(10): 1435-1442
[65] Kumaresh S, Tejraj M, J. Control. Release, 2001, 70:1-20
[66] Christine Allen a , Dusica, Colloids and Surfaces B: Biointerfaces ,1999, 16:
3-27
[67] 常津，刘海峰，许晓秋，中国生物医学工程学报，2002， 12
[68] 邵健，杨宇民，南通医学学报，1999
[69] Lingyun Chen, Yunmin Du, Xiaoqing Zeng, Carbohydrate Research，2003, 338：
333-340
[70] 陈洁，夏文水，日用化学工业，1999
105
参考文献 参考文献
[71] 罗彦凤，王远亮，高分子材料科学与工程，2003,19
[72] 何永言，夏淑贞，吴蒙等，高分子材料科学与工程，1993，2：24-28.
[73] 沈兴海 高宏成，化学通报，1995
[74] Sanjeeb K.Sahoo,Jayanth Panyam,et al,Journal of Controlled Release,
[75] L D Richard ,J O Hollinger eds.CRC Press, Inc.boca Raton.,1995: 17-31
[76] Gliding D K,Reed A M., Journal of Polymer,1979,20:1459
[77]VacantiJ.P.,MayerJ.E,Langer R.,et al.,Journal of annthorac
surg,1995,60:513-517
[78] Lexroux J.C.,Allamann E, Journal of Controlled Release,1996,39:339
[79] 赵锋，高永良，中国新药杂志，2002, 11
[80] 崔正刚，殷福珊，中国轻工业出版社，1999
[81] Lagourette B,Peyrelasse J,Boned C,et al., Nature,1979,281:60-62.
[82] 李晓，刘戈，袁惠根等，福州大学学报，2000，8
[83] 施利毅，张潘琪，高分子材料科学与工程，1999，7
[84] Gabriele Reich， Journal of Pharmaceutics and Biopharmaceutics， 1998，45：
165-171
[85] 赵锋，高永良，中国新药杂志，2002，11
[86] Schakenraad J.M.,Hardon M.J.,Feijen J.,et al., Journal of biomedical Materials
Research,1990,24:529-545.
[87] Bodmeier R.,Chen H., International Journal of Pharmartics,1989,51:1-8
[88] Vert M,Mauduit J,Li S.,biomaterials,1994,15:1209-1213
[89] Spenlehauer G,Vert M,Benoit J.P.,et,al.,biomaterials,1989,10:557-563
[90] Isobe M.,Yamazaki Y., Oida S-I., et al., Journal of biomedical Materials
Research, 1996,32:433-438
[91] Kamei S,Inoue Y., Advanced Polymer Science,1999,94:107-141
[92] James M.Anderson,Matthew S.Shive, Advanced Drug Delivery Reviews, 1997,
28:5-24
[93] 中华人民共和国药典二部（2000 版），北京，中国标准出版社，2000：472～474
[94] 沈正荣，朱家蕙，中国医药工业杂志，1996，28(7):306～308
[95] M.L.Hans,A.M.Lowman, 2002: 319-327
[96] Friederike von Burkersroda,Luise Schedl,Biomaterials,2002,23: 4221-4231
[97] 杨炳兴，成国祥，胶体与聚合物，2003
[98] L.B, Peppas,Inter. J. Pharm., 1995, 116:1～9
[99] Kerr J.R.,Harmon B.V.,Cancer,1994,73:2013
106
参考文献 参考文献
[100] Crystal R.G.,Science,1995,270～404
[101] Schafield J.P.,Caukey C.T.,Br Med Bull,1995,51(1):56～71
[102] Yamamoto T.,Kamata N.,Kawano H.,et al.,Cancer Res., 1986,44:414-416
[103] Michael F. Haller, Maryland， 2001
[104] 黄世文，卓仁禧，科学通报 48 卷 第五期,2003
[105] Cohen.L.S.,J.E.Rosenthal,D.W.Horner,American Journal of Cardiology
29:520-523
[106] Agrawal.C.M.and K.A.Athanasiou, J.of Biomed. Material. Research，1997，
38(2):105-114
[107] Eldridge.J.H.,C.J.Hammond, J. of Control. Release, 2:205-214
[108] Yamamoto T., Kamata N, Kawano H et al, Cancer Res., 1986,44:41-416
[109] Liberman T.A., Nusbaum H.R., Razon N., et al, Nature, 1985, 313: 14-147
[110] Luo D Saltzman M.,Synthetic,Natural Biotechnology,2000,18:33-37
[111] Nakanishi M.,Akuta T,Nagoshi E,et al.,European Journal Pharmartics
Science,2001.13:17-24
[112] Rolland A P.,Drug Delivery,1998,15:143-198
[113] Ferber B.,Science,2001,294:1638-1642
[114] Son K,Huang L., Proc Natl Acad Sci USA,1994,91:12669-12672
[115] Alexander IE, Russell DW, Miller AD., J Virol, 1994, 68:8282-8287
[116] Kuriyama S, Masui K, Sakamoto T, et al, Anticancer Res. 1998, 18:3399-3406.
[117] 陈刚 李世拥 于波，中华胃肠外科杂志，2002，1（5）：134－136