溴芬酸钠／壳聚糖纳米粒蛋白胶胶联羊膜的制备及其对兔角膜新生血管的影响  

Preparation of bromfenac sodium/chitosan nanoparticles loaded fibrin-binding amniotic membrane and its effect on rabbit corneal neovascularization in vitro

作　　者：邬桂玉[1] 赵敏[1] 计岩[1] 

机构地区：[1]重庆医科大学附属第一医院眼科,重庆市眼科学重点实验室,重庆400016

出　　处：《第三军医大学学报》2013年第6期495-499,共5页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2008AC0086);重庆医科大学附属第一医院课题(YXJJ 2009-09)~~

摘　　要：目的制备一种具有双重缓释功能的新型生物复合材料溴芬酸钠/壳聚糖纳米粒胶联羊膜(BF/CS-FBAM),初步研究其体外缓释性能及对角膜新生血管的影响。方法采用离子胶联法制备溴芬酸钠/壳聚糖纳米粒并观察其表征,将载药纳米粒混入纤维蛋白胶中,再将混合物喷涂到羊膜表面制成BF/CS-FBAM,并进行形态学和体外释药观察,最后通过兔角膜碱烧伤模型观察BF/CS-FBAM对角膜新生血管的影响,用实时荧光定量PCR方法检测术后角膜中VEGF的表达。结果溴芬酸钠/壳聚糖纳米粒呈球形,表面光滑,大小均一,其粒径为(548.7±14.4)nm,Zeta电位为(29.8±2.2)mV,包封率为(67.11±4.56)%,载药量为(57.71±5.38)%。载有纳米药的纤维蛋白胶与羊膜黏合紧密,表面呈网状结构,纳米药充斥其中。在体外,BF/CS-FBAM的释药可达14 d。在碱烧伤模型中,术后BF/CS-FBAM组和BF/CS组在减少炎症反应、抑制新生血管及降低VEGF表达方面都显著优于对照组,且各时间点的差异有统计学意义(P<0.05);同时,术后14 d和21 d BF/CS-FBAM组中VEGF的表达明显低于BF/CS组(P<0.05)。结论制备的BF/CS-FBAM作为一种新型生物移植材料可在局部缓慢释放溴芬酸钠,并对角膜新生血管的产生有较好的抑制性。Objective To prepare a novel biological composite material, bromfenac sodium/chitosan nanopartiele loaded fibrin-binding amniotie membrane （BF/CS-FBAM）, with double sustained-released function, and investigate the drug release in vitro and its effect on rabbit corneal neovascularization （CNV）. Methods The ionotropic gelation was used to produce bromfenac sodium/ehitosan nanoparticles （BF/CS）. After its physic properties were identified, the BF/CS nanoparticles were mixed with fibrin glue （FG） , and then sprayed to the surface of amniotic membrane （AM）. After BF/CS-FBAM was obtained, the morphological changes and drug release in vitro were observed and measured. The effect of BF/CS-FBAM for CNV was observed in rabbit corneal alkali burn model, meanwhile the expression of vascular endothelial growth factor （VEGF） in the cornea were quantified by reverse transcription-quantitative PCR （RT-qPCR）. Results The nanoparticles were smooth and spherical, in a diameter of 548.7 ＋ 14.4 nm, and had a positive zeta potential of 29. 8 ＋ 2.2 mV. The encapsulation efficiency and loading capacity of BF/CS nanoparticles were （ 67. 11 ＋ 4. 56）% and （57.71 _＋ 5.38）% respectively. The nanoparticle-loaded fibrin gel was firmly adhered to the AM. Moreover, nanoparticles were coated within FBAM symmetrically, and its surface had a netted structure. The BF released from the BF/CS-FBAM was sustained for approximately 14 d in vitro. In rabbit corneal alkali burn model, BF/CS-FBAM group and BF/CS group had better curative effect in attenuating inflammatory reaction, anti-CNV and down-regulating VEGF than control group in every time point after injury （P 〈 0. 05 ）. Meanwhile, the expression of VEGF in BF/CS-FBAM group was significantly lower than BF/CS group in the 14th and 21st day after injury （P 〈 0. 05 ）. Conclusion As a novel biological implantable material, the fabricated BF/CS-FBAM locally releases BF in a sustained way and inhibits the generation of CNV.

关 键 词：溴芬酸钠 纤维蛋白胶 壳聚糖 纳米粒 羊膜 角膜新生血管 

分 类 号：R318.08[医药卫生—生物医学工程] R772.2[医药卫生—基础医学]