Angiopep-2修饰核-壳介孔二氧化硅脂质囊纳米粒的制备及体外评价

Preparation and in Vitro Evaluation of Paclitaxel-loaded Core-Shell Structural Phospholipid-Functionalized Mesoporous Silica Nanoparticles Modified with Angiopep-2

目的制备angiopep-2修饰核-壳结构介孔二氧化硅脂质囊纳米粒（ANG-MSN-LP），并进行体外评价。方法采用Stober法制备介孔二氧化硅纳米粒（MSN），改进了溶液吸附法来制备载紫杉醇（PTX）的介孔二氧化硅纳米粒（MSN-PTX），然后以载紫杉醇的介孔二氧化硅纳米粒为核，运用自组装和薄膜水化法构建核-壳结构的angiopep-2修饰介孔二氧化硅脂质囊纳米粒（ANG-MSN-LP-PTX），透析袋法考察体外释药特性，噻唑蓝（MTT）法考察载体对人脑微毛细血管内皮细胞（HBMEC）和C6的细胞毒性，建立体外血脑屏障（BBB）模型研究载体对紫杉醇的跨膜转运能力和对C6细胞周期的影响。结果介孔二氧化硅纳米粒粒径为（94．61±3．91）nm[PDI（0．085±0．01）]，比表面积（SBET）为425m2·g^-1，孔容积（y。）为0．37cm3·g^-1，孔径3．5nm。介孔二氧化硅纳米粒在紫杉醇的饱和溶液中反复吸附7次时达到平衡，载药量高达11．1％。Angiopep-2修饰介孔二氧化硅脂质囊纳米粒分布均一，无团聚现象，具有明显的核．壳结构，粒径（106．37±3．76）nm[PDI（0．14±0．02）]，并且在0～10μg·mL^-1范围内对HBMEC和C6细胞具有良好的安全性和生物相容性。Angiopep-2修饰介孔二氧化硅脂质囊纳米粒体外释药48h内的累积释放率达到75．5％，突释现象降低，具有明显的缓释特性；跨膜血脑屏障转运12h后，对紫杉醇转运率高达10．74％；在细胞周期实验中，使c6细胞滞留在G2-M期细胞数比例为（40．92±6．20）％，显著高于各对照组数倍。结论Angiopep-2修饰介孔二氧化硅脂质囊纳米粒是一种优良的递药载体并有望应用于脑胶质瘤的治疗；反复饱和溶液吸附法可有效提高药物载药量，值得应用和借鉴。

OBJECTIVE To prepare and evaluate the novel core-shell structural phospholipid-functionalized mesoporous silica nanoparticles （MSN-LP） modified with angiopep-2 （ANG-MSN-LP）. METHODS Mesoporous silica nanoparticles （MSN） was syn- thesized by the modified Stober method. MSN-PTX was prepared by saturated solution adsorption method. ANG-MSN-LP was developed by selfassembly and film hydration method. By using dialysis bag method to investigate the in vitro drug release characteristics and MTT method to investigate the cytotoxicity on HBMEC and C6 cells. The transport ability and effects on cell cycle of the carrier was investi- gated by the BBB monolayer model. RESULTS MSN was synthesized with high specific surface area （ SBrT, 425 m2 ·g-1 ）, cumula- tive pore volume （ Vp, 0. 37 cm3 · g-1 ） and pore size（3.5 nm）. PTX was highly encapsulated （drug loading efficiency up to 11.1% ） into MSN. Results of in vitro release showed that about 75.5% of FFX released from ANG-MSN-LP-PTX after 48 h and burst release was effectively reduced compared with MSN-PTX or PTX solution, indicating pronounced sustained-release characteristics. The good biocompatibility and low toxicity of ANG-MSN-LP were evaluated by HBMEC and C6 cells. The transport ratio was 2.49% for PTX, 2. 72% for MSN-PTX, 4.45% for MSN-LP-PTX and 10. 74% for ANG-MSN-LP-PTX respectively. In addition, ANG-MSN-LP-PTX showed a higher cell number of G2-M phase of 40. 92± 6. 20%. CONCLUSION ANG-MSN-LP is a prospective targeting drug deliv- ery system for therapy of brain glioma. Meanwhile, saturated solution adsorption method can increase the drug loading efficiency highly.