加替沙星-壳聚糖纳米粒胶联羊膜药膜的制备及体外实验研究

Preparation and in Vitro Study of Gatifloxacin-loaded Chitosan Nanoparticle-impregnated Fibrin Glue-binding Amniotic Membrane

目的制备一种新型生物多功能复合材料——加替沙星-壳聚糖纳米粒胶联羊膜药膜,并观察其体外缓释性能及抑菌作用。方法采用离子胶联法制备加替沙星-壳聚糖纳米粒并检测其表征,然后将载药纳米粒加入到纤维蛋白胶中,混匀后再与羊膜胶联制成加替沙星-壳聚糖纳米粒胶联羊膜药膜,光镜及扫描电镜下观察其形态学特征,高效液相色谱法测定其中加替沙星的浓度,微量肉汤稀释法检测其体外抑菌活性。结果加替沙星纳米粒的粒径为291±33 nm,载药量为10.46%,包封率为61.32%。载有纳米药物的纤维蛋白胶与羊膜基底面粘合紧密,呈圆形的加替沙星纳米粒附着在胶原纤维的网状结构上。在体外,加替沙星-壳聚糖纳米粒胶联羊膜药膜可持续释药达15天,对金黄色葡萄球菌和铜绿假单胞菌的最低抑菌浓度分别为0.05 μg/ml和0.42μg/ml。结论加替沙星-壳聚糖纳米粒胶联羊膜药膜在体外具有良好的缓释性能及抑菌作用。

Objective To develop a novel biological multi-functional composite materials--Gatifloxacin-loaded chitosan nanopardcle-impregnated fibrin glue-binding amniotie membrane （GAT-CS-FGAM） and study its drug release characteristics and antibacterial activity in vitro. Methods A Gatifloxacin-loaded chitosan nanopartielc （GAT-CS） was prepared by Ionotropic gelification and its character were assayed. GAT-CS was added into fibrin glue （FG） , and then sprayed it to the entire basal surface of amniotic membrane （AM） after enough immingle;a GAT-CS-FGAM was prepared. The morphology of GAT-CS-FGAM was observed with Light Microscope and Scanning Electron Microscope,its concentration of gatifloxacin was detected by high performance liquid chromatography, and the nfinimum inhibitory concentrations （ MIC ） was evaluated using broth microdilution antifungal susceptibility test. Results The mean size of the GAT-CS was 291 ± 33 nm,loading capacity was 10.46%, and encapsulation efficiency was 61.32%. The nanoparticle-impregnated fibrin glue could firmly cohere with the amniotic membrane,and the round nanoparticles adhere to the reticular network of collagenous fibers. In vitro, the drug release of GAT-CS- FGAM was very slow,and can sustain for 15 days. The MIC of GAT-CS-FGAM against Staphylococcus aureus and Pseudomonas aeruglnosa were O. 05 μg/ml and 0. 42 μg/ml,respeetively. Conclusions The GAT-CS-FGAM in vitro take on a perfect sustained release property and antibacterial activities.