5-氟脲嘧啶壳聚糖缓释微球抑制裸鼠腹腔种植胃癌细胞生长的实验研究
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摘要
目的:
探讨5-氟脲嘧啶壳聚糖缓释微球对裸鼠腹腔种植胃癌细胞生长的抑制作用。
方法:
采用乳化化学交联法制备5-氟脲嘧啶壳聚糖缓释微球:将5-氟脲嘧啶溶于5%醋酸溶液,加入壳聚糖,配成水相;以石蜡油为油相;将水相滴入油相,超声乳化, 25%戊二醛交联固化,真空干燥得微球。同法,仅不加5-氟脲嘧啶制得空白壳聚糖微球。观察微球的基本性状,并测定5-氟脲嘧啶壳聚糖缓释微球的药物包封率及载药量。将微球142 mg(含5-氟脲嘧啶50mg)及5-氟脲嘧啶粉剂50mg分别置于透析袋中,每小时1次,24h后每天1次,共7d,测定各时段微球释药量及药物累计释放率。建立裸鼠胃癌细胞腹腔种植模型,按体表面积计算5-氟脲嘧啶用量,换算出5-氟脲嘧啶壳聚糖缓释微球用量,将祼鼠随机分为4组:A组(5-氟脲嘧啶壳聚糖缓释微球组)、B组(空白壳聚糖微球组)、C组(5-氟脲嘧啶组)、D组(生理盐水组),分别于裸鼠腹腔内注入5-氟脲嘧啶壳聚糖缓释微球、空白壳聚糖微球、5-氟脲嘧啶注射剂、生理盐水。全部裸鼠在SPF条件下饲养至1组裸鼠全部死亡。观察各组祼鼠的体重、腹围变化、腹腔癌结节计数、腹水胃癌细胞凋亡率、祼鼠生存率和药物毒副作用。
结果:
所制5-氟脲嘧啶壳聚糖缓释微球分布均匀,平均粒径为(185.5±15.0)nm,载药量为(35.2±0.8)%,药物包封率为(49.3±2.1)%。体外药物缓释实验显示,所制5-氟脲嘧啶壳聚糖缓释微球第1h 5-氟脲嘧啶释放量为21.4±2.0μg/ml,以后逐渐减少,第1d 5-氟脲嘧啶累计释放量为123.2±3.6μg/ml,以后呈持续缓慢释放,至第7d仍释放4.5±0.6μg /ml。在4组裸鼠中,A组裸鼠的腹围、体重在用药前后无明显改变(P >0.05),A组裸鼠的生存率明显高于B组裸鼠(P<0.01)、C组裸鼠(P<0.05)和D组裸鼠(P<0.01),并且,A组裸鼠腹腔癌结节数也较其余3组裸鼠明显减少(P <0.05),而且,腹水胃癌细胞凋亡率明显高于B组祼鼠(P<0.05)、C组祼鼠(P<0.05)和D组祼鼠(P<0.01);然而, A组裸鼠静脉血WBC总数、ALT水平、BUN水平、TBI水平与其他三组裸鼠比较无明显差异(P> 0.05)。
结论:
5-氟脲嘧啶壳聚糖缓释微球粒径分布均匀,具有较好的药物包封率及药物缓释性;用于胃癌腹腔化疗可明显抑制祼鼠腹腔种植胃癌细胞的生长,改善胃癌细胞腹腔种植祼鼠的预后。
Objective: Investigation the growth inhibitory effection of 5- fluorouracil loaded chi- tosan sustained-release microspheres to the gastric cancer cells of peritoneal implantation in nude mice.
Methods: The microspheres were prepared using emulsified-chemical cross-linking method : the water phase which consisted of 5-fluorouracil dissolved in 5% acetic acid and chitosan added after 5-fluorouracil was dissolved was droppded into the oil phase of paraffin oil , emulsification by ultrasound and using 25% glutaraldehyde to cross-link, then obtained the microspheres by vacuum drying. The same method, only without 5 - fluorouracil obtained blank microspheres. Observed basic characteristics of the microspheres and studied the drug encapsulation efficiency and the Drug load- ing. Weighed the microspheres 142 mg (include 5 - fluorouracil 50mg)and 5 - fluorouracil powder 50mg , placed them in dialysis bags, observed the role of drug release in vitro by determining con- centration and cumulative release hourly in 24h、then Once a day and a total of seven days . Establish the model of peritoneal implantation of gastric cancer cells in nude mice, calculated according to body surface area of 5 - fluorouracil dosage, con- version out of the dosage of 5- fluorouracil loaded chitosan sustained-release micro- spheres. All 32 nude mice divided into four groups randomly: A group(5- fluorouracil loaded chito- san sustained-release microspheres group)、B group(blank microspheres group)、C group(5- fluorouracil group)and D group(NS group), the nude mice were in- jected into the abdominal cavity 5- fluorouracil loaded chitosan sustained-release microspheres、blank microspheres、5- fluorouracil、NS by groups. Studied the changes of the every mouse body weight and abdominal circumference、the number of peritoneal metastatic noduli、the rate of gastric cancer cells apoptosis、survival of the nude mouse and the drug toxicity.
Results: The average particle size of the microspheres was (185.5±15.0)nm, drug-loading rate was (35.2±0.8)% and encapsulation efficiency was ( 49.3±2.1)%. The microspheres release experiments in vitro showed that it released 5-fluorouracil 21.4±3.6μg/ml in the first hour、123.2±3.6μg/ml on the first day and 4.5±0.6/ml on the seventh day by releasing slowly. In the nude mice of A group ,The survival rate was more than the nude mice of B group (P< 0.05)、the nude mice of C group(P< 0.05)and the nude mice of D group(P< 0.01), the number of peritoneal metastatic noduli in the nude mice of A group was less than the nude mice of other group(P<0.05), the rate of gastric cancer cells apoptosis in the nude mice of A group was more than the nude mice of B group (P< 0.05)、the nude mice of C group(P<0.05)and the nude mice of D group(P< 0.01) when the difference in the number of WBC、ALT、BUN、TBI of Vein blood in the nude mice compared with other groups showed no statistical significance (P> 0.05).
Conclusions: The 5- fluorouracil loaded chitosan sustained- release microspheres have uniform distribution、good drug encapsulation efficiency and drug release. The microspheres show perfect growth inhibition effect to gastric cancer cells of peritoneal implantation in nude mice and can improve the prognosis of the mice.
探讨5-氟脲嘧啶壳聚糖缓释微球对裸鼠腹腔种植胃癌细胞生长的抑制作用。
方法:
采用乳化化学交联法制备5-氟脲嘧啶壳聚糖缓释微球:将5-氟脲嘧啶溶于5%醋酸溶液,加入壳聚糖,配成水相;以石蜡油为油相;将水相滴入油相,超声乳化, 25%戊二醛交联固化,真空干燥得微球。同法,仅不加5-氟脲嘧啶制得空白壳聚糖微球。观察微球的基本性状,并测定5-氟脲嘧啶壳聚糖缓释微球的药物包封率及载药量。将微球142 mg(含5-氟脲嘧啶50mg)及5-氟脲嘧啶粉剂50mg分别置于透析袋中,每小时1次,24h后每天1次,共7d,测定各时段微球释药量及药物累计释放率。建立裸鼠胃癌细胞腹腔种植模型,按体表面积计算5-氟脲嘧啶用量,换算出5-氟脲嘧啶壳聚糖缓释微球用量,将祼鼠随机分为4组:A组(5-氟脲嘧啶壳聚糖缓释微球组)、B组(空白壳聚糖微球组)、C组(5-氟脲嘧啶组)、D组(生理盐水组),分别于裸鼠腹腔内注入5-氟脲嘧啶壳聚糖缓释微球、空白壳聚糖微球、5-氟脲嘧啶注射剂、生理盐水。全部裸鼠在SPF条件下饲养至1组裸鼠全部死亡。观察各组祼鼠的体重、腹围变化、腹腔癌结节计数、腹水胃癌细胞凋亡率、祼鼠生存率和药物毒副作用。
结果:
所制5-氟脲嘧啶壳聚糖缓释微球分布均匀,平均粒径为(185.5±15.0)nm,载药量为(35.2±0.8)%,药物包封率为(49.3±2.1)%。体外药物缓释实验显示,所制5-氟脲嘧啶壳聚糖缓释微球第1h 5-氟脲嘧啶释放量为21.4±2.0μg/ml,以后逐渐减少,第1d 5-氟脲嘧啶累计释放量为123.2±3.6μg/ml,以后呈持续缓慢释放,至第7d仍释放4.5±0.6μg /ml。在4组裸鼠中,A组裸鼠的腹围、体重在用药前后无明显改变(P >0.05),A组裸鼠的生存率明显高于B组裸鼠(P<0.01)、C组裸鼠(P<0.05)和D组裸鼠(P<0.01),并且,A组裸鼠腹腔癌结节数也较其余3组裸鼠明显减少(P <0.05),而且,腹水胃癌细胞凋亡率明显高于B组祼鼠(P<0.05)、C组祼鼠(P<0.05)和D组祼鼠(P<0.01);然而, A组裸鼠静脉血WBC总数、ALT水平、BUN水平、TBI水平与其他三组裸鼠比较无明显差异(P> 0.05)。
结论:
5-氟脲嘧啶壳聚糖缓释微球粒径分布均匀,具有较好的药物包封率及药物缓释性;用于胃癌腹腔化疗可明显抑制祼鼠腹腔种植胃癌细胞的生长,改善胃癌细胞腹腔种植祼鼠的预后。
Objective: Investigation the growth inhibitory effection of 5- fluorouracil loaded chi- tosan sustained-release microspheres to the gastric cancer cells of peritoneal implantation in nude mice.
Methods: The microspheres were prepared using emulsified-chemical cross-linking method : the water phase which consisted of 5-fluorouracil dissolved in 5% acetic acid and chitosan added after 5-fluorouracil was dissolved was droppded into the oil phase of paraffin oil , emulsification by ultrasound and using 25% glutaraldehyde to cross-link, then obtained the microspheres by vacuum drying. The same method, only without 5 - fluorouracil obtained blank microspheres. Observed basic characteristics of the microspheres and studied the drug encapsulation efficiency and the Drug load- ing. Weighed the microspheres 142 mg (include 5 - fluorouracil 50mg)and 5 - fluorouracil powder 50mg , placed them in dialysis bags, observed the role of drug release in vitro by determining con- centration and cumulative release hourly in 24h、then Once a day and a total of seven days . Establish the model of peritoneal implantation of gastric cancer cells in nude mice, calculated according to body surface area of 5 - fluorouracil dosage, con- version out of the dosage of 5- fluorouracil loaded chitosan sustained-release micro- spheres. All 32 nude mice divided into four groups randomly: A group(5- fluorouracil loaded chito- san sustained-release microspheres group)、B group(blank microspheres group)、C group(5- fluorouracil group)and D group(NS group), the nude mice were in- jected into the abdominal cavity 5- fluorouracil loaded chitosan sustained-release microspheres、blank microspheres、5- fluorouracil、NS by groups. Studied the changes of the every mouse body weight and abdominal circumference、the number of peritoneal metastatic noduli、the rate of gastric cancer cells apoptosis、survival of the nude mouse and the drug toxicity.
Results: The average particle size of the microspheres was (185.5±15.0)nm, drug-loading rate was (35.2±0.8)% and encapsulation efficiency was ( 49.3±2.1)%. The microspheres release experiments in vitro showed that it released 5-fluorouracil 21.4±3.6μg/ml in the first hour、123.2±3.6μg/ml on the first day and 4.5±0.6/ml on the seventh day by releasing slowly. In the nude mice of A group ,The survival rate was more than the nude mice of B group (P< 0.05)、the nude mice of C group(P< 0.05)and the nude mice of D group(P< 0.01), the number of peritoneal metastatic noduli in the nude mice of A group was less than the nude mice of other group(P<0.05), the rate of gastric cancer cells apoptosis in the nude mice of A group was more than the nude mice of B group (P< 0.05)、the nude mice of C group(P<0.05)and the nude mice of D group(P< 0.01) when the difference in the number of WBC、ALT、BUN、TBI of Vein blood in the nude mice compared with other groups showed no statistical significance (P> 0.05).
Conclusions: The 5- fluorouracil loaded chitosan sustained- release microspheres have uniform distribution、good drug encapsulation efficiency and drug release. The microspheres show perfect growth inhibition effect to gastric cancer cells of peritoneal implantation in nude mice and can improve the prognosis of the mice.
引文
[1] Parkin DM,Pisani P,Ferlay J.Estimates of the worldwide incidence of 25 major cancers in 1991[J].Int J Cancer,1999,80(2):827-841.
[2] Yoo CH ,Noh SH, et a1.Recurrence following curative resection for gastric carcinoma[J].Br J Surg, 2000,87(2):236-242.
[3]陆枫林,陆亚琴.腹腔化疗前后对腹水端粒酶活性影响研究[J].中华消化杂志, 2004,2(2):110-111.
[4] Elias D,Bonnay M,Puizillou JM,et a1.Heated intraoperative intraperitoneal oxaliplatin after complete resection of peritoneal carcinomatosis:pharmacokinetics and tissue distribut- ion[J].Ann OncoL, 2002,13(2):267-272.
[5] Shanmuganathan S, Shanumugasundaram N, Adhirajan N,et al. Prepara- tion and characterization of chitosan microspheres for doxycycline delivery[J].Cabohydr Polym,2008, 73(2):201-211.
[6] Park SH,Chun MK,Choi HK.Preparation of an extended-release martrix tablet using chitonsan/ Carbopol interpolyer conplex[J].Int J Pham, 2008,347(1-2):39-44.
[7]蒋智清,林友文,陈兰妹.氟脲嘧啶微球体外释药的影响因素[J].福建医科大学学报,2006,40(7):364-368.
[8] Kim T H,Park I K,Nah J W ,et a1.Galactosylated chitosan/ DNA nanoparticles prepared using water-soluble chitosan as a gene car- rier[J].Biomaterials,2004,25 (3):3783-3792.
[9]徐叔云,卞如濂,陈修,等.药理实验方法学[M].第三版.北京:人民卫生版社,2002,565.
[10]李年丰.腹腔内化疗对胃肠恶性肿瘤的治疗[J].中国现代医学杂志,2005,15(7):1023-1026.
[11] Tsujitani S,Fukuda K,Saito H,et a1.Administration in a hypotonic intraperitoneal eisplatin during operation as treatment for peritoneal dissemination of gastric cancer[J].Surgery,2002,13(1):98-104.
[12] Roth AD,Ajani J. Docetavel-based chemotherapy in the treatment of gastric cancer[J].Ann Oncol,2003,14(4):1141-1144.
[13]陈小春,潘凯,张桂英,等.胃癌患者术中腹腔内温热灌注化疗的临床研究[J].中华胃肠外科杂志,2004.7(7):134-136.
[14]张阳德,吴泽建,龚连生.半乳糖化白蛋白磁性阿霉素纳米粒在正常大鼠体内的分布[J].中国医学工程,2004,12(4):5-7.
[15]郑爱萍,刘海宏,李宏斌,等.5-氟脲嘧啶壳聚糖微球的制备及体外释放特性[J].中国药科大学学报,2004,35(4):318-323.
[16] Vila A,Sa,nchez A,Janes K,et al.Low molecular weight chitosan nanoparticles as new carriers for nasal vaccine delivery in mice [J] . Europ Journal of Pharmaceutics Biopharmaceutics,2004,57(2):123-131.
[17]张洪,黄徐英.三七总皂苷壳聚糖缓释微球的制备及体外释放特性研究[J].广东药学院学报, 2006,22(5):54-56.
[18] Xu YM.Du YM.Effect of molecular structure of chitosan on protein delivery properties of ehitosan nanoparticles[J].Int J Pharm,2003,250(10):215.
[19]李和平,肖华伍,龙姝,等.5-氟脲嘧啶壳聚糖缓释微球的制备[J].长沙理工大学学报(自然科学版),2006,3(4):91-97.
[20]卢剑,常津,赵硕,等.载药纳米微粒表面性能与表面改性[J].化学通报,2008,66(6):50-56.
[21] Hagiwara A ,Takahashi T ,Yamagishi H. Activated carbon particles adsorbing MMC used against peritoneal carcinomatosis in gastric cancer[M] . Springer ,Tokyo. 1999,45:214-217.
[22] Lv Xiaoying,SU Miancheng,LI You,el a1.Efect of Acanthopanax giraldii Harms Var.Hispidus Hoo polysaccharides on the human gastric cancer cell line SGC-7901 and its possible mechanism[J]. Chin Med J,2002,115(5):716-721.
[1] Vila A,Sanchez A,Janes K,et al.Low molecular weight chitosan nanoparticles as new carriers for nasal vaccine delivery in mice[J].Europ J Pharm Biopharm,2004,57(7):123-131.
[2] Kim T H,Park I K,Nah J W ,et al.Galactosylated chitosan/ DNA nanoparticles prepared using water-soluble chitosan as a gene carrier[J].Biomatefials,2004,25 (7):3783-3792.
[3] Alpar H O,Somavarapu s,Atuah K N,et a1.Biodegradable mucoadhesive particulates for nasal and pulmonary antigen and DNA delivery[J].Adv Drug Deliv Rev,2005,57(3): 411430.
[4] Rokhade A P,Shelke N B,Patil S A,et a1.Novel interpenetrating ploymer network micospheres of chitosan and methylcellulose for controlled release of theophylline [J].Carbohydr polym,2007,69(4):678-687.
[5]郑连英,朱江峰,孙昆山.壳聚糖的抗菌性能研究[J].材料科学与工程,2000,18(2):22-24.
[6] [6]王红昌,孙晓飞.不同分子量高脱乙酰度壳聚糖的制备及表征[J].中国海洋药物杂志,2007,26(1):16-19.
[7] MUZZARELLI R A A,TARAI O,FILIPPINI E,et a1.Antimicrobial Properties of N-carboxylbutyl Chitosan[J].Antimicro Ag Chemotherap,1990,34(2):2019-2023.
[8] Seyfar th F, SchliemannS, E sner P, et al . Antifungal effect of high-and low-molecular -weight chitosanhydr ochl oride, carbo- xymethyl chitosan, chitosan oli gosaccharide and Nacetyl-D- glucosamine against Candidaal bicans, Candi dakrusei and Candi daglabrata[J] . Int J Pharm, 2008, 353(1-2):139-148.
[9]陈威,吴清平,张菊梅,等.壳聚糖抑菌机制的初步研究[J] .微生物学报. 2008,48(2):164-168.
[10]宋永焕,刘一,于铁成,等.壳聚糖纳米颗粒免疫刺激作用的初步研究[J].浙江医学,2008,50(8):1208-1200.
[11] Cho E J, Rahman MA, Kim S W,et al . Chitosan oli gosaccharides inhibit adi pogenesis in 3T3-L1 adi pocytes[J] . J Microbi ol Biotechnol , 2008,18(1): 80-87.
[12]房国坚,蒋玉燕,毕忆群,等.壳聚糖纳米混悬剂抗突变生物学效应:原核细胞基因突变试验[J].中国组织工程研究与临床,2008,12(4):8867-8869.
[13] Shanmuganathan S,Shanumugasundaram N,Adhirajan N,et al.Preparation and characterization of chitosan microspheres for doxycycline delivery[J].Cabohydr Polym,2008,73(2):201-211.
[14] Park SH,Chun MK,Choi HK.Preparation of an extended-release martrix tablet using chitonsan/ Carbopol interpolyer conplex[J].Int J Pham, 2008,347(1-2):39-44.
[15] Janes KA.Fresneau M P,Marazuela A,et al.Chitosan nanoparticles as delivery systems fordoxorubicin[J].J Control Release,2001;73(3):255-267.
[16]王知非,李贺书.纳米技术在肿瘤防治中的应用[J].实用肿瘤学杂志,2002,16(1):75-79.
[17]王健,冯玉杰.壳聚糖磁性微球的制备和工艺参数的优化[J].磁性材料及器件,2008,39(6):44-46.
[18] Bodmeier R,OH K H,Pramar Y.Preparation and evaluation of drug- containiing chitosan beads[J].Dmg Dev.Ind.Pharm,1989,15(2):1475.
[19] Bodmeier R,OH K H,Pramar Y.Preparation and evaluation of drug-containiing chitosan beads[J].Dmg Dev.Ind.Pharm,1989, 15:1475.
[20] Jamela S R,Kumary T V,Lal A V.Progesterone-loaded chitosn microspheres:a long acting biodegradable controlled delivery system[J].Control Rel,1998,52(1-2):17.
[21] Pan Y,Li YJ,Zhao HY,et a1.Bioadhesive polysaceharide in protein delivery system :chitosan nanoparticles improve the intestinal absorption of insulin in vivo[J].Int J Pharm ,2002,249(5):l39.
[22] Fernandez-Urrusuno R,Cairo P.Remunan-Lopez C,et al.Enhancement of nasal absorption of insulin using chitosan nanoparticles [J].Pharm Res,1999,16(10):1589.
[23] Janes KA,Fresneau MP,Maraznela A,et a1.Chitosan nanoparticles as delivery systems for doxorubicin[J].J Controlled Release,2001,73(3):255.
[24] Ohya Y,Shiratani M,Kobayashi H,et a1.Release behavior of 5-fluorouraeil from chitosan-gel nanospheres immobilizing 5-fluorouracil coated with polysaccharides and their cell specific cytotoxicity[J].Pure Appl Chem,1994,31(1):629-642.
[25] Tanima B,Susmita M,Singh K,et a1.Preparation,characterizalion and biodistribution of ultrafine chitosan nanoparlicles[J].Int J Pharm ,2002,243(1-2):93-102.
[26] Berthold A,Cremer K,Kreuter J.Preparation and characterizalion of chitosan microspheres as drug carrier for prednisolone sodium phosphate as model for anti-inflammatory drugs[J].J Cont Rel,1996,39(1):17-25.
[27]刘利萍,李苹,吴泽志,等.5-FU壳聚糖/丝素复合磁微球的制备及体外性质研究[J].中国药学杂志,2003,38(1O):774-777.
[28] Chellat N , Tabrizian M , Dumitriu S , et a1 . Study of biodegradation behavior of chitosan-xanthan microspheres in simulated physio- logical media[J].J Biomed Mater Res,2OO0,53(5):592-599.
[29] Lamprecht A,Yamamoto H,Takeuehi H,et a1.pH-semitive microsphere delivery increases oral bioavailability of calcitonin[J].Con- trolled Release,2004,98(1):1-9.
[30] Filipovie-Grcie J,Perissutti B,Moneghini M,et a1.Spraydriedcarbamazepine loaded chitosan and HPMC microspheres:preparation and haracterisation[J].Pharra Pharmacol,2003,55(7):921-931.
[31] Zhengrong.Cui,Russell J.Mumper,chitosan-based nanopartides for topical genetic immunization[J].J Cont Rel,2001,75(3):409-4l9.
[32]昝青峰,成鹏,李兆新等.壳聚糖交联特性对壳聚糖/β-TCP微球成球性的影响[J].稀有金属材料与工程,2008,36(12):54-57.
[33] Su Young Chae,Sohee Son,Minhyung Lee,el a1.Deoxycholic acid-conjugaled chitosan oligosaccharide nanoparticles for efficient gene carrier[J].J Cont Rel,2005,109(1- 3):33O-344.
[34] Bugamelli F,Raggi MA,Orienti I,el a1.Controlled insulin release from chitosan microparticles[J].Arch pharm,1998,33 1(4):l33-l38.
[35] Mooren FC,Berlhold A,Domschke W,et a1.Influence of chitosan microspheres on the transport of prednisolone sodium phosphate across HT-29 cell monolayers[J].Fharm Res,1998;15(1):58-65.
[36] Hwang HY, Ki mI S, Kwon I C, et al . Tumor target ability and antitumor effect of docetaxel - loaded hydrophobically modified glycol chitosan nanoparticles[J] . J Control Release, 2008, 128(1) : 23- 31.
[37] Kim J H, Kim YS, Park K, et al . Antitumor efficacy of cisplatin- loaded glycol chitosan nanoparticles in tumor-bearing mice[J].J Control Release, 2008, 127(1): 41- 49.
[38] Min KH, Park K, Kim YS, et al . Hydrophobically modified glycol chitosan nanoparticles-encapsulate dcanp to thecin enhance the drug stability and tumor targeting in cancer therapy[J].J Control Release, 2008, 127(3):208- 218.
[39] Alexakis T,Boadi DK,Quong D,et a1.Microencapsulation of DNA within alginate microspheres and crosslinked chitosan membranes for in vivo application[J] . Appl Biovechnol,1995,50(1):93-1O6.
[40] de la Fuente M, Seijo B, Al onso MJ . Bioadhesi ve hyaluronan-chitosan nanoparticles can transport genes across the ocular mucosa and transfect ocular tissue[J] . Gene Ther , 2008, 15( 9) : 668- 676.
[41] Khatri K, Goyal AK, Gupta P N, et al . Plasmid DNA loaded chitosan nanoparticles for nasal mucosal immunization against hepatitis B[J] . Int J Pharm, 2008, 354(1- 2) : 235- 241.
[42] Jameela SR,Latha PG,Subramamoniam A,et a1.Antitumour activity of mitoxantrone-loaded chitosan microspheres against Ehrlich ascites carcinoma[J].J Pharm Pharmacol,1996;48:685.
[43]王亚玲,黄玲玲,邵健.壳聚糖结肠靶向释药体系的初步研究[J].交通医学,2008,22(6):595-597.
[44] Grenha A, Remu C, Carvalho E L, et al .Microspheres containing lipid/ chitosan nanopar ticles complexes for pulmonary delivery of therapeutic proteins[J] .Eur J Pharm Biopharm, 2008, 69(1) : 83-93.
[45]贾栋,高国栋,李永林.壳聚糖纳米粒载体转染铜超氧化物歧化酶对缺血再灌注脑组织的作用[J].中国组织工程研究与临床, 2008,12(45):8827- 8830.
[2] Yoo CH ,Noh SH, et a1.Recurrence following curative resection for gastric carcinoma[J].Br J Surg, 2000,87(2):236-242.
[3]陆枫林,陆亚琴.腹腔化疗前后对腹水端粒酶活性影响研究[J].中华消化杂志, 2004,2(2):110-111.
[4] Elias D,Bonnay M,Puizillou JM,et a1.Heated intraoperative intraperitoneal oxaliplatin after complete resection of peritoneal carcinomatosis:pharmacokinetics and tissue distribut- ion[J].Ann OncoL, 2002,13(2):267-272.
[5] Shanmuganathan S, Shanumugasundaram N, Adhirajan N,et al. Prepara- tion and characterization of chitosan microspheres for doxycycline delivery[J].Cabohydr Polym,2008, 73(2):201-211.
[6] Park SH,Chun MK,Choi HK.Preparation of an extended-release martrix tablet using chitonsan/ Carbopol interpolyer conplex[J].Int J Pham, 2008,347(1-2):39-44.
[7]蒋智清,林友文,陈兰妹.氟脲嘧啶微球体外释药的影响因素[J].福建医科大学学报,2006,40(7):364-368.
[8] Kim T H,Park I K,Nah J W ,et a1.Galactosylated chitosan/ DNA nanoparticles prepared using water-soluble chitosan as a gene car- rier[J].Biomaterials,2004,25 (3):3783-3792.
[9]徐叔云,卞如濂,陈修,等.药理实验方法学[M].第三版.北京:人民卫生版社,2002,565.
[10]李年丰.腹腔内化疗对胃肠恶性肿瘤的治疗[J].中国现代医学杂志,2005,15(7):1023-1026.
[11] Tsujitani S,Fukuda K,Saito H,et a1.Administration in a hypotonic intraperitoneal eisplatin during operation as treatment for peritoneal dissemination of gastric cancer[J].Surgery,2002,13(1):98-104.
[12] Roth AD,Ajani J. Docetavel-based chemotherapy in the treatment of gastric cancer[J].Ann Oncol,2003,14(4):1141-1144.
[13]陈小春,潘凯,张桂英,等.胃癌患者术中腹腔内温热灌注化疗的临床研究[J].中华胃肠外科杂志,2004.7(7):134-136.
[14]张阳德,吴泽建,龚连生.半乳糖化白蛋白磁性阿霉素纳米粒在正常大鼠体内的分布[J].中国医学工程,2004,12(4):5-7.
[15]郑爱萍,刘海宏,李宏斌,等.5-氟脲嘧啶壳聚糖微球的制备及体外释放特性[J].中国药科大学学报,2004,35(4):318-323.
[16] Vila A,Sa,nchez A,Janes K,et al.Low molecular weight chitosan nanoparticles as new carriers for nasal vaccine delivery in mice [J] . Europ Journal of Pharmaceutics Biopharmaceutics,2004,57(2):123-131.
[17]张洪,黄徐英.三七总皂苷壳聚糖缓释微球的制备及体外释放特性研究[J].广东药学院学报, 2006,22(5):54-56.
[18] Xu YM.Du YM.Effect of molecular structure of chitosan on protein delivery properties of ehitosan nanoparticles[J].Int J Pharm,2003,250(10):215.
[19]李和平,肖华伍,龙姝,等.5-氟脲嘧啶壳聚糖缓释微球的制备[J].长沙理工大学学报(自然科学版),2006,3(4):91-97.
[20]卢剑,常津,赵硕,等.载药纳米微粒表面性能与表面改性[J].化学通报,2008,66(6):50-56.
[21] Hagiwara A ,Takahashi T ,Yamagishi H. Activated carbon particles adsorbing MMC used against peritoneal carcinomatosis in gastric cancer[M] . Springer ,Tokyo. 1999,45:214-217.
[22] Lv Xiaoying,SU Miancheng,LI You,el a1.Efect of Acanthopanax giraldii Harms Var.Hispidus Hoo polysaccharides on the human gastric cancer cell line SGC-7901 and its possible mechanism[J]. Chin Med J,2002,115(5):716-721.
[1] Vila A,Sanchez A,Janes K,et al.Low molecular weight chitosan nanoparticles as new carriers for nasal vaccine delivery in mice[J].Europ J Pharm Biopharm,2004,57(7):123-131.
[2] Kim T H,Park I K,Nah J W ,et al.Galactosylated chitosan/ DNA nanoparticles prepared using water-soluble chitosan as a gene carrier[J].Biomatefials,2004,25 (7):3783-3792.
[3] Alpar H O,Somavarapu s,Atuah K N,et a1.Biodegradable mucoadhesive particulates for nasal and pulmonary antigen and DNA delivery[J].Adv Drug Deliv Rev,2005,57(3): 411430.
[4] Rokhade A P,Shelke N B,Patil S A,et a1.Novel interpenetrating ploymer network micospheres of chitosan and methylcellulose for controlled release of theophylline [J].Carbohydr polym,2007,69(4):678-687.
[5]郑连英,朱江峰,孙昆山.壳聚糖的抗菌性能研究[J].材料科学与工程,2000,18(2):22-24.
[6] [6]王红昌,孙晓飞.不同分子量高脱乙酰度壳聚糖的制备及表征[J].中国海洋药物杂志,2007,26(1):16-19.
[7] MUZZARELLI R A A,TARAI O,FILIPPINI E,et a1.Antimicrobial Properties of N-carboxylbutyl Chitosan[J].Antimicro Ag Chemotherap,1990,34(2):2019-2023.
[8] Seyfar th F, SchliemannS, E sner P, et al . Antifungal effect of high-and low-molecular -weight chitosanhydr ochl oride, carbo- xymethyl chitosan, chitosan oli gosaccharide and Nacetyl-D- glucosamine against Candidaal bicans, Candi dakrusei and Candi daglabrata[J] . Int J Pharm, 2008, 353(1-2):139-148.
[9]陈威,吴清平,张菊梅,等.壳聚糖抑菌机制的初步研究[J] .微生物学报. 2008,48(2):164-168.
[10]宋永焕,刘一,于铁成,等.壳聚糖纳米颗粒免疫刺激作用的初步研究[J].浙江医学,2008,50(8):1208-1200.
[11] Cho E J, Rahman MA, Kim S W,et al . Chitosan oli gosaccharides inhibit adi pogenesis in 3T3-L1 adi pocytes[J] . J Microbi ol Biotechnol , 2008,18(1): 80-87.
[12]房国坚,蒋玉燕,毕忆群,等.壳聚糖纳米混悬剂抗突变生物学效应:原核细胞基因突变试验[J].中国组织工程研究与临床,2008,12(4):8867-8869.
[13] Shanmuganathan S,Shanumugasundaram N,Adhirajan N,et al.Preparation and characterization of chitosan microspheres for doxycycline delivery[J].Cabohydr Polym,2008,73(2):201-211.
[14] Park SH,Chun MK,Choi HK.Preparation of an extended-release martrix tablet using chitonsan/ Carbopol interpolyer conplex[J].Int J Pham, 2008,347(1-2):39-44.
[15] Janes KA.Fresneau M P,Marazuela A,et al.Chitosan nanoparticles as delivery systems fordoxorubicin[J].J Control Release,2001;73(3):255-267.
[16]王知非,李贺书.纳米技术在肿瘤防治中的应用[J].实用肿瘤学杂志,2002,16(1):75-79.
[17]王健,冯玉杰.壳聚糖磁性微球的制备和工艺参数的优化[J].磁性材料及器件,2008,39(6):44-46.
[18] Bodmeier R,OH K H,Pramar Y.Preparation and evaluation of drug- containiing chitosan beads[J].Dmg Dev.Ind.Pharm,1989,15(2):1475.
[19] Bodmeier R,OH K H,Pramar Y.Preparation and evaluation of drug-containiing chitosan beads[J].Dmg Dev.Ind.Pharm,1989, 15:1475.
[20] Jamela S R,Kumary T V,Lal A V.Progesterone-loaded chitosn microspheres:a long acting biodegradable controlled delivery system[J].Control Rel,1998,52(1-2):17.
[21] Pan Y,Li YJ,Zhao HY,et a1.Bioadhesive polysaceharide in protein delivery system :chitosan nanoparticles improve the intestinal absorption of insulin in vivo[J].Int J Pharm ,2002,249(5):l39.
[22] Fernandez-Urrusuno R,Cairo P.Remunan-Lopez C,et al.Enhancement of nasal absorption of insulin using chitosan nanoparticles [J].Pharm Res,1999,16(10):1589.
[23] Janes KA,Fresneau MP,Maraznela A,et a1.Chitosan nanoparticles as delivery systems for doxorubicin[J].J Controlled Release,2001,73(3):255.
[24] Ohya Y,Shiratani M,Kobayashi H,et a1.Release behavior of 5-fluorouraeil from chitosan-gel nanospheres immobilizing 5-fluorouracil coated with polysaccharides and their cell specific cytotoxicity[J].Pure Appl Chem,1994,31(1):629-642.
[25] Tanima B,Susmita M,Singh K,et a1.Preparation,characterizalion and biodistribution of ultrafine chitosan nanoparlicles[J].Int J Pharm ,2002,243(1-2):93-102.
[26] Berthold A,Cremer K,Kreuter J.Preparation and characterizalion of chitosan microspheres as drug carrier for prednisolone sodium phosphate as model for anti-inflammatory drugs[J].J Cont Rel,1996,39(1):17-25.
[27]刘利萍,李苹,吴泽志,等.5-FU壳聚糖/丝素复合磁微球的制备及体外性质研究[J].中国药学杂志,2003,38(1O):774-777.
[28] Chellat N , Tabrizian M , Dumitriu S , et a1 . Study of biodegradation behavior of chitosan-xanthan microspheres in simulated physio- logical media[J].J Biomed Mater Res,2OO0,53(5):592-599.
[29] Lamprecht A,Yamamoto H,Takeuehi H,et a1.pH-semitive microsphere delivery increases oral bioavailability of calcitonin[J].Con- trolled Release,2004,98(1):1-9.
[30] Filipovie-Grcie J,Perissutti B,Moneghini M,et a1.Spraydriedcarbamazepine loaded chitosan and HPMC microspheres:preparation and haracterisation[J].Pharra Pharmacol,2003,55(7):921-931.
[31] Zhengrong.Cui,Russell J.Mumper,chitosan-based nanopartides for topical genetic immunization[J].J Cont Rel,2001,75(3):409-4l9.
[32]昝青峰,成鹏,李兆新等.壳聚糖交联特性对壳聚糖/β-TCP微球成球性的影响[J].稀有金属材料与工程,2008,36(12):54-57.
[33] Su Young Chae,Sohee Son,Minhyung Lee,el a1.Deoxycholic acid-conjugaled chitosan oligosaccharide nanoparticles for efficient gene carrier[J].J Cont Rel,2005,109(1- 3):33O-344.
[34] Bugamelli F,Raggi MA,Orienti I,el a1.Controlled insulin release from chitosan microparticles[J].Arch pharm,1998,33 1(4):l33-l38.
[35] Mooren FC,Berlhold A,Domschke W,et a1.Influence of chitosan microspheres on the transport of prednisolone sodium phosphate across HT-29 cell monolayers[J].Fharm Res,1998;15(1):58-65.
[36] Hwang HY, Ki mI S, Kwon I C, et al . Tumor target ability and antitumor effect of docetaxel - loaded hydrophobically modified glycol chitosan nanoparticles[J] . J Control Release, 2008, 128(1) : 23- 31.
[37] Kim J H, Kim YS, Park K, et al . Antitumor efficacy of cisplatin- loaded glycol chitosan nanoparticles in tumor-bearing mice[J].J Control Release, 2008, 127(1): 41- 49.
[38] Min KH, Park K, Kim YS, et al . Hydrophobically modified glycol chitosan nanoparticles-encapsulate dcanp to thecin enhance the drug stability and tumor targeting in cancer therapy[J].J Control Release, 2008, 127(3):208- 218.
[39] Alexakis T,Boadi DK,Quong D,et a1.Microencapsulation of DNA within alginate microspheres and crosslinked chitosan membranes for in vivo application[J] . Appl Biovechnol,1995,50(1):93-1O6.
[40] de la Fuente M, Seijo B, Al onso MJ . Bioadhesi ve hyaluronan-chitosan nanoparticles can transport genes across the ocular mucosa and transfect ocular tissue[J] . Gene Ther , 2008, 15( 9) : 668- 676.
[41] Khatri K, Goyal AK, Gupta P N, et al . Plasmid DNA loaded chitosan nanoparticles for nasal mucosal immunization against hepatitis B[J] . Int J Pharm, 2008, 354(1- 2) : 235- 241.
[42] Jameela SR,Latha PG,Subramamoniam A,et a1.Antitumour activity of mitoxantrone-loaded chitosan microspheres against Ehrlich ascites carcinoma[J].J Pharm Pharmacol,1996;48:685.
[43]王亚玲,黄玲玲,邵健.壳聚糖结肠靶向释药体系的初步研究[J].交通医学,2008,22(6):595-597.
[44] Grenha A, Remu C, Carvalho E L, et al .Microspheres containing lipid/ chitosan nanopar ticles complexes for pulmonary delivery of therapeutic proteins[J] .Eur J Pharm Biopharm, 2008, 69(1) : 83-93.
[45]贾栋,高国栋,李永林.壳聚糖纳米粒载体转染铜超氧化物歧化酶对缺血再灌注脑组织的作用[J].中国组织工程研究与临床, 2008,12(45):8827- 8830.