载microRNA-148b壳聚糖/透明质酸纳米粒制备及鉴定

Preparation and characterization of chitosan/hyaluronic acid nanoparticles for microRNA-148b delivery

目的:制备并评价壳聚糖/透明质酸(CS/HA)纳米粒负载microRNA-148b(miR-148b)的效果。方法:制备CS/HA/miR-148b纳米粒,用激光纳米测量仪和透射电镜对其粒径、电位和形态进行观察。通过凝胶阻滞实验观察CS/HA纳米粒对miR-148b的包载情况。用CCK-8实验评价CS/HA/miR-148b纳米粒对大鼠骨髓间充质干细胞(MSCs)的细胞毒性;转染实验和流式细胞术检测转染效率。结果:CS/HA/miR·148b纳米粒呈球形,粒径为160~370 nm,电位为+15 mV^+41 mV。氮磷比为20可以达到最佳的包封效果。CS/HA/miR-148b纳米粒对大鼠MSCs无细胞毒性并且具有较高的转染效率。结论:CS/HA纳米粒可以安全有效地将miR-148b转染入MSCs中。

Objective： To prepare and characterize chitosarr/hyaluronic acid （CS/HA） nanoparticles for miRNA-148b（miR-148b） delivery. Methods： CS/HA/miR-148b nanoparticles were prepared. The particle size, zeta potential, morphology were investigated by nano laser granulometer and TEM respectively. The encapsulation efficiency of CS/HA nanoparticles was evaluated by gel retarding analysis. The cytotoxicity and transfection efficiency of CS/HA/miR-148b nanoparticles on rat bone marrow mesenchymal stem cells （MSCs） were evaluated by CCK-8 assay, transfection assay and FCM respectively. Results： The CS/HA/miR-148b nanoparticles were discrete spherical particles with the diameter of 160 to 370 nm and zeta potential of ＋ 15 to ＋41 mV. N/P ratio at 20：1 of the nanoparticles showed the optimum encapsulation efficiency for miR-148b. CS/HA/miR-148b nanoparticles exhibited no cytotoxicity to MSCs and could deliver miR-148b to the MSCs at high transfection efficiency. Condusion： CS/HA/miR-148b nanoparticles are safe and effective for miR-148b delivery into MSCs.