5-氟尿嘧啶纳米药膜的体外缓释及其对结肠癌细胞抑制作用研究

5-FU Loaded Nanofibers Membrane for Controlled Release Effect Against Colon Cancer Cell in Vitro

目的研究担载抗肿瘤药物5-氟尿嘧啶(5-FU)的左旋聚乳酸(PLLA)共混羊毛蛋白纳米纤维药膜的缓释功能及其对结肠癌细胞的体外抑制效果。方法将PLLA与羊毛蛋白共混交联,并加入抗肿瘤药物5-FU,利用高压静电纺丝技术将共混溶液制作成具有缓释效果的纳米纤维薄膜。通过扫描电镜(SEM)观察其形貌。以高效液相色谱法(HPLC)测定5-FU/PLLA羊毛蛋白药膜中5-FU的缓释效果。采用噻唑蓝(MTT)比色法,检测药膜对结肠癌细胞HCT116的体外杀伤效果。同时采用倒置相差显微镜、透射电子显微镜(TEM)观测实验组细胞的生长情况。结果由SEM可以看出5-FU能够均匀分布在纳米纤维药膜中,HPLC显示5-FU能够缓慢释放并基本符合零级动力学规律。采用不同处理因素后,通过显微镜观察,药膜浸泡时间越长的实验组细胞出现肿胀、凋亡、坏死的情况越明显。MTT检测纳米药膜实验组的细胞存活率为(47.5±2.8)%,而不含药物的纯膜组对细胞基本无影响,其存活率为(93.9±2.8)%,差异具有统计学意义(P<0.01)。结论 5-FU/PLLA羊毛蛋白纳米药膜有明显缓释效果,具有较好的肿瘤细胞抑制作用和潜在的医疗应用价值。

Objective To investigate the controlled release effect and the anticancer cell ability of a 5FU loaded polyLlactic acid （PLLA） nanofibers membrane blending with keratin. Methods Making PLLA and keratin mix together and crosslinking to generate blending solution. Then the anticancer drug 5FU was added into the solution to fabricate nanofibers membrane by high voltage electrospinning method. The micro morphology was observed by scanning electron microscope （SEM）. The controlled release effect of 5FU from the nanofibers membrane was measured by high performance liquid chromatography （HPLC）. The eytotoxicity of 5FU/PLLA keratin nanofibers membrane was evaluated by using 3（4, 5dimethylthiazol2yl） 2, 5diphenyltetrazolium bromide （MTT） assay on HCT116 cell lines. At the meantime, cell growth morphology of HCT116 in experiment group were observed by microscope and transmission electron microscope. Results 5FU could be dispersed homogeneous in the PLLA/keratin nanofibers membrane through SEM. HPLC suggested that 5FU could be diffused out from the fibers slowly and uniformly, which corresponded the zero order kinetics basically. After different treatment, the longer time the 5FU/PLLA keratin nanofibers （experiment group） immerse in the medium, the much more swelling, apoptosis, and necrocytosis of the cells were observed. The cell viability for experiment group was （47.5±2.8） % by MTT, while the PLLA keratin nanofibers without 5FU had no significant impact on cell viability （93.9±2.8） %, which was statistic significance （P 〈 0. 01）.Conclusion 5FU/PLLA keratin nanofibers membrane owns good controlled release effect and satisfies cell inhibitory effect against HCT116 cells in vitro, which suggested that it has a promising prospect for clinical therapy.