5-Fu纳米毫微粒的制备及体外对晶状体上皮细胞增生的抑制作用

Preparation and in vitro inhibitive effect in lens epithelial cell proliferation of 5-fluorouracil-loaded nanoparticles

目的制备5氟尿嘧啶(5fluorouracil,5Fu)聚乳酸纳米毫微粒并观察其表征及体外对兔晶状体上皮细胞的抑制作用。方法采用复乳法制备5Fu聚乳酸纳米毫微粒,观察毫微粒的大小、表面形态和结构,毫微粒载药率和体外药物释放曲线。取传3代晶状体上皮细胞进行对比试验,设5Fu纳米毫微粒、5Fu原药、空载纳米毫微粒组、空白对照组。设0.5～62.5mg·L-14个剂量组,作用24h、72h、120h、168h后光镜下观察细胞性状,MTT法检测抑制作用。结果(1)制备的5Fu纳米毫微粒平均粒径(191.000±0.202)nm;载药率为8.1%;首日药物突释率为38.3%,缓释时间20d;(2)5Fu纳米毫微粒和5Fu原药对兔晶状体上皮细胞均有抑制作用,并随时间和剂量的增加而增加。空载聚乳酸纳米毫微粒对细胞无抑制作用;(3)作用初期各剂量组5Fu纳米毫微粒的抑制作用低于或等于原药,随时间延长分别在不同时间点超过原药,差异有显著性。结论5Fu聚乳酸纳米毫微粒性状稳定,可长期释放药物,有效抑制兔晶状体上皮细胞的增生,随时间延长作用明显高于原药,可以作为靶向缓释药物载体。

Objective To prepare and observe the character of 5-fluoronracil-loaded polyactic acid nanoparticles [ PLA（ 5-Fu）-NP ], and to study the inhibitive effect in lens epithelial cell proliferation tn vitro. Methods PLA（5-Fn）-NP were prepared by a double emulsion technique, and the particles were characterized for size, modality, encapsulation efficiency,and tn vttro release. The third passage lens epithelial cells were divided into four groups：PLA（5-Fu）-NP,original 5-Fu,blank nanoparticles and control. Four doses of 5-Fu were tested：0.5mg·L^-1,2.5mg2L^-1,12.5mg·L^-1,62.5mg·L^-1.After 24h,72 h, 120 h, 168 h, the cells growth conditions were observed with light microscope and evaluated by MTT colorimetry. Results PLA（5-Fu）-NP had an average diameter of about （ 191. 000 ± 0. 202） nm and a drug loading of 8. 1%. After a fast release （about 38.3%） during the first day, a more gradual drug release was sustained about 20 days. Both PLA（5-Fu）-NP and original 5-Fu could inhibit the proliferation of lens epithelial cells in dose-dependent and time-dependent manner. The inhibitive effect of PLA（5-Fu）-NP was more effective than original 5-Fu after a period of time. Blank nanoparticles had no cytotoxicity on lens epithelial cells. Conclusion PLA （5-Fu）-NP can be proposed as a potential controlled and targeted ophthalmic delivery system for the treatment of posterior capsular opacification.