聚乙烯亚胺修饰的荧光素钠纳米颗粒在荧光素眼底血管造影术的应用和安全性  

Applicationand safetyof polyethyleneimine modified fluorescein sodium nanoparticles in fundus fluorescein angiography

作　　者：杨倩 蔡雯婷[2] 金惠子[2] 余咚卉 沈天怡 于靖 YANG Qian;CAI Wenting;JIN Huizi;YU Donghui;SHEN Tianyi;YU Jing(Shanghai Clinical College,Anhui Medical University,Hefei 230032,Anhui Province,China;Department of Ophthalmology,Shanghai Tenth People’s Hospital Affiliated to Tongji University,Shanghai 200072,China)

机构地区：[1]安徽医科大学上海临床学院,安徽省合肥市230032 [2]同济大学附属第十人民医院眼科,上海市200072

出　　处：《眼科新进展》2020年第11期1005-1010,共6页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号81470648);中央高校基本科研业务费专项资金(编号22120180509)。

摘　　要：目的探讨聚乙烯亚胺修饰的荧光素钠(PEI-NHAc-FS)纳米颗粒(nanoparticles,NPs)在荧光素眼底血管造影术的应用和安全性。方法选取90只健康棕色雄性挪威大鼠作为实验动物。将1 mL荧光素钠(200 g·L^-1)加入10 mol的1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDC)混合物中搅拌30 min,再加入10 mol的N-羟基琥珀酰亚胺搅拌3 h。然后将该溶液加入到5 mL聚乙烯亚胺(polyethyleneimine,PEI)(76 g·L^-1)中,剧烈搅拌3 d获得PEI-NH2-FS。将2 mL三乙胺加入到PEI-NH2-FS原液中,30 min后再将1 mL的乙酸酐加入混合物中搅拌24 h,合成PEI-NHAc-FS。使用核磁共振波谱仪和紫外可见光吸收光谱观察PEI-NHAc-FS的结构,并对合成的NPs结构进行表征;CCK-8检测其细胞毒性。选取30只大鼠,根据荧光素钠和PEI-NHAc-FS NPs的浓度(5 g·L^-1、10 g·L^-1、50 g·L^-1)将大鼠分为6组,每组5只,分别于注射前及注射后5 min、20 min、30 min和60 min进行眼底摄像,采集大鼠眼底血管荧光素图像。用激光诱导大鼠脉络膜新生血管模型后,取10只大鼠,随机分为10 g·L^-1荧光素钠组和PEI-NHAc-FS NPs组,每组5只,分别于注射前及注射后5 min、20 min、30 min和60 min通过眼底血管造影成像观察病灶处渗漏。另取40只大鼠,随机分为10 g·L^-1荧光素钠组和PEI-NHAc-FS NPs组,每组20只,于注射后10 min、20 min、30 min和60 min进行荧光冰冻切片。将10只大鼠随机分为对照组(注射生理盐水)和PEI-NHAc-FS NPs组,每组5只,行组织切片HE染色和视网膜电图检测。采用HE染色和视网膜电图观察PEI-NHAc-FS NPs在体内的生物安全性。结果通过核磁共振和紫外可见光吸收光谱观察到荧光素钠与PEI成功耦合。发射荧光光谱显示,游离荧光素钠和PEI-NHAc-FS NPs的最大发射波长分别出现在546 nm和544 nm处。CCK-8检测结果表明,不同浓度的荧光素钠和PEI-NHAc-FS NPs处理人脐静脉内皮细胞12 h、24 h后,细胞活性差异无统计学�Objective To investigatethe application and safety of fluorescein sodium(PEI-NHAc-FS)nanoparticles(NPs)modified with polyethyleneimine in fundus fluorescein angiography(FFA).Methods Ninety healthy brown male brown Norway rats were used as experimental animals.We took 1 mL fluorescein sodium(200 g·L^-1)in 10 mol of 1-ethyl-3-(3-dimethylaminopropy)carbodiimide hydrochloride mixture and stirred for 30 minutes,followed by the addition of 10 mol equivalents of the N-hydroxysuccinimide and stirring for another 3 hours.Next,this solution was added into the 5 mL PEI(76 g·L^-1)under vigorous stirring for 3 days to obtain PEI-NH2-FS.Then the solution was added to 5 mL PEI(76 g·L^-1),and PEI-NH2-FS was obtained by vigorous stirring for 3 days.Briefly,2 mL triethylamine was added to the raw solution of the PEI-NH2-FS.After 30 minutes,1 mL acetic anhydride to the remaining primary amines in PEI was added to the mixture and stirred for 24 hours and PEI-NHAc-FS was synthesized.Nuclear magnetic resonance(NMR)spectrometerand ultraviolet-visible spectroscopy was recorded to observe the structure of PEI-NHAc-FS and performed to characterize(NPs).The CCK-8 was used to evaluate the cell cytotoxicity.According to the concentrations of fluorescein sodium and PEI-NHAc-FS NPs(5 g·L^-1,10 g·L^-1,50 g·L^-1),thirty rats were randomly divided into 6 groups(5 rats in each group)for fundus angiography before and 5 minutes,20 minutes,30 minutes and 60 minutes after injection.Laser-induced choroidal neovascularization model was established.Forty rats in 90 rats were randomly divided into 10 g·L^-1 fluorescein sodium group and PEI-NHAc-FS NPs group(20 rats in each group),followed by fluorescence frozen section imaging 10 minutes,20 minutes,30 minutes and 60 minutes after injection.Ten rats in 90 rats were randomly divided into control group(normal saline was given)and PEI-NHAc-FS NPs group(5 rats in each group).HE staining and electroretinogram were used to observe the biosafety of PEI-NHAc-FS NPs in vivo.Results The successful coupling of

关 键 词：荧光素钠 荧光素眼底血管造影术 聚乙烯亚胺 造影剂 

分 类 号：R770.4[医药卫生—眼科]