载汉防己甲素壳聚糖纳米微球的制备及其对人翼状胬肉细胞增殖的抑制作用

目的　通过制备新型载汉防己甲素的壳聚糖纳米微球，研究其对于人翼状胬肉细胞增殖的抑制作用。方法　化学合成新型壳聚糖衍生物 （deoxycholic acid-modified chitosan，DAMC），与汉防己甲素作用合成载药纳米微球，并检测其性能。载药纳米微球作用体外培养的人翼状胬肉成纤维细胞第1天、3天、5天后，采用CCK-8法检测人翼状胬肉成纤维细胞的活性。结果　通过化学反应成功获得脱氧胆酸基团接枝的DAMC，可包载汉防己甲素药物，两者形成的载药纳米微球载药量较高，可高达76%。粒径50~500 nm，Zeta电位始终保持正电位。体外药物释放实验显示载药纳米微球缓释汉防己甲素可达48 h。细胞活性实验显示Tet组第1天、3天、5天细胞活性分别为（60.70±2.30）%、（50.22±2.35）%、（21.99±2.07）%，而Tet/DAMC组分别为（79.77±2.09）%、（63.24±2.83）%、（40.28±1.19）%，含10×10^-6 mol·L^-1汉防己甲素的载药纳米微球具有抑制人翼状胬肉成纤维细胞的作用，且细胞毒性较原药明显降低。结论　载药纳米微球具有缓释药物的作用，对人翼状胬肉成纤维细胞的增殖具有明显的抑制作用，且细胞毒性较原药明显降低，有望提高汉防己甲素防治翼状胬肉复发的效果。

Preparation of chitosan nano-particles loaded tetrandrine and its inhibitory effects on the proliferation of human pterygium fibroblast cells

Objective　To investigate the inhibitory effects of chitosan nano-particles loaded tetrandrine（Tet） on proliferation of cultured pterygium fibroblasts.Methods　The deoxycholic acid-modified chitosan （DAMC） derivative was synthesized through amidation reaction，and their properties were investigated.The viability of human pterygium fibroblasts （HPF） was evaluated by cell counting kit-8(CCK-8) assay after cells were interacted with Tet/DAMC nano-particles on day 1，3 and 5.Results　The synthesized chitosan derivative and Tet formed drug-loaded nano-particles，and the agent-loading capacity was approximately 76%，and the sizes of agent-loaded nano-particles were 50-500 nm，with Zeta potential values being positive.The result of in vitro drug release experiment indicated that the nano-particles constantly released Tet in a controlled manner within 48 h.The viability of HPF in Tet group was (60.70±2.30)%，(50.22±2.35)%，(21.99±2.07)% on day 1，3，5，respectively，but the corresponding data in Tet/DAMC group was (79.77±2.09)%，(63.24±2.83)%，(40.28±1.19)%，respectively.The CCK-8 assay demonstrated that the Tet/DAMC nano-particles could inhibit HPF proliferation，and presented lower toxicity than Tet alone.Conclusion　Chitosan nano-particles loaded tetrandrine exhibits a sustained agent-release behavior，which has obvious inhibitory effects on the proliferation of human pterygium fibroblasts，but its cytotoxicity is significantly lower than the original drug’s，thereby possessing a great promise for improving the outcome of Tet for the prevention of pterygium recurrence.