低分子量鱼精蛋白修饰的PLGA纳米粒应用于内耳毛细胞递送的分布特性考察

Investigation on the distribution characteristics of PLGA nanoparticles modified with low molecular weight protamine delivered into inner ear hair cells

目的对低分子量鱼精蛋白(low molecular weight protamine,LMWP)修饰的PLGA纳米粒在内耳毛细胞的分布特性进行考察,旨在为防治内耳疾病的药物递送提供新载体。方法利用LMWP对PLGA纳米粒进行修饰,并进行表征;通过HEI-OC1细胞模型考察纳米粒的细胞毒性;采用激光扫描共聚焦荧光显微镜技术对纳米粒进入斑马鱼的能力进行评价,同时考察纳米粒在豚鼠耳蜗和基底膜的分布情况。结果 LMWP修饰PLGA纳米粒的分散性良好,粒径分布均匀;纳米粒具有较低的细胞毒性,且能够更有效地被斑马鱼侧线毛细胞摄取。与未修饰的纳米粒相比,LMWP修饰显著增强纳米粒在豚鼠耳蜗的分布,并且明显促进基底膜中回和顶回毛细胞的摄取。结论 LMWP修饰的PLGA纳米粒具有良好的细胞相容性,能有效递送至斑马鱼和豚鼠毛细胞,有望成为内耳给药的新型载体。

Objective To investigate the distribution characteristics of low molecular weight protamine(LMWP)-modified PLGA nanoparticles in inner ear hair cells,in order to provide a novel carrier for drug delivery in the treatment of inner ear diseases.Methods The nanoparticles were modified by LMWP and characterized.The cytotoxicity of the NPs was evaluated by HEI-OC1 cell model.The ability of NPs entering zebrafish was evaluated by Laser-scanning confocal microscopy(LSCM),and the distribution of NPs in guinea pigs was investigated by whole body imaging system and LSCM.Results The results of size distribution and TEM image showed the nanoparticles had a spherical and relative uniform size.The NPs had low cytotoxicity and could be more effectively absorbed by zebrafish lateral hair cells.Compared with the unmodified NPs,LMWP-modified PLGA NPs significantly enhanced the distribution of NPs in the cochlea,and significantly promoted the uptake of NPs in the middle and apical turns of basilar membrane.Conclusion LMWP-modified PLGA nanoparticles have good cytocompatibility and can be effectively distributed to zebrafish lateral hair cells,as well as inner hair cells of guinea pigs,which is expected to become a new carrier for drug delivery in inner ear.