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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 study the sustained-release function of PLLA blended wool protein nanofibers loaded with 5-fluorouracil (5-FU) and its inhibitory effect on colon cancer cells in vitro. Methods PLLA and wool protein were blended and cross-linked, and the antitumor drug 5-FU was added. The blend solution was made into a nanofiber membrane with sustained release effect by high-pressure electrospinning. Morphology was observed by scanning electron microscopy (SEM). The sustained-release effect of 5-FU in 5-FU / PLLA wool protein membrane was determined by high performance liquid chromatography (HPLC). MTT assay was used to detect the cytotoxicity of the drug on colon cancer cell line HCT116 in vitro. At the same time, the phase contrast microscope and transmission electron microscope (TEM) were used to observe the growth of the experimental group cells. Results SEM showed that 5-FU could be uniformly distributed in the nanofiber membrane. HPLC showed that 5-FU could release slowly and basically met the zero order kinetics. After using different treatment factors, through the microscope observation, the longer the experimental group, the membrane swelling, apoptosis, necrosis of the more obvious. MTT assay showed that the survival rate of the experimental group was (47.5 ± 2.8)%, while the drug-free pure membrane group had no effect on the cells, the survival rate was (93.9 ± 2.8)%, the difference was statistically significant ( P <0.01). Conclusion The 5-FU / PLLA wool protein nano-membrane has a significant sustained release effect, with good tumor cell inhibition and potential medical value.