胶原/纤维蛋白胶复合药膜的性质及其体外抑瘤效应初步研究

Characteristic of collagen/fibrin complex film and the preliminary study of its in vitro anti-tumor effect

目的 制备载硫酸长春新碱（VCR）微球的胶原/纤维蛋白胶的缓释药膜并研究其性质,考察京尼平的交联浓度（0、0.05、0.1、0.2 mol/ml）对药膜性质的影响.方法 冷冻干燥和京尼平交联法制备多孔胶原膜,将VCR微球混合纤维蛋白胶后加入多孔胶原膜中制备胶原/纤维蛋白胶复合药膜,考察药膜的表面形态、机械性能、降解性质和体外释药行为,并用CCK-8法检测药膜对3种肿瘤细胞（人肺癌细胞株A549、人乳腺癌细胞株MCF-7和人宫颈癌细胞株HeLa）的体外抑瘤率.结果 随着交联剂京尼平浓度的增加,药膜机械强度增加,而断裂伸长率降低.VCR微球与纤维蛋白胶及胶原复合制备成药膜,可达到双重缓释的作用,减少药物突释,并延缓药物释放.未交联的F0药膜药物24h突释为（41.8±3.4）％,京尼平交联后的F0.05、F0.1和F0.2药膜的药物突释分别为（38.4±4.1）％、（35.2±3.6）％和（34.3±3.7）％.未载药的F0.1药膜无显著细胞毒效应,而载药的F0.1药膜在不同时间释放的药液对3种肿瘤细胞株均有显著的抑制作用,且对各细胞株敏感度不同.结论 F0.1药膜能不同程度减少药物的突释,使药物释放更加平稳缓慢,有望用作抗肿瘤药物载体.

Objective To investigate the properties of vincristine sulfate （VCR） loaded collagen/fibrin microspheres complex film,especially to give evaluation of the concentration of genipin （0,0.05,0.1 and 0.2 mol/ml） as a cross-linking agent on the properties of the film.Methods Porous collagen film were fabricated by freeze-drying method and cross-linked by genipin.VCR loaded microspheres were mixed with fibrin into collagen scaffolds to fabricate the complex film.The physiochemical properties of the film,such as surface morphology,mechanical function,in vitro degradation and VCR release kinetics were also measured.In vitro cytotoxicities of VCR loaded film were evaluated using CCK-8 assay.Results The tensile strength of the film was strengthened and the breaking elongating rate of the film decreased with more proportion of genipin.The degradation rates of the films （F0.05,F0.1 and F0.2） were slower than that of the F0 film.VCR was released from the film in a prolonged period and the initial burst release of the film was less significant.The initial release of F0 was （41.8±3.4）%,while of the films which cross linked （F0.05,F0.1 and F0.2） were （38.4±4.1）%,（35.2±3.6）% and （34.3±3.7）%,respectively.The results showed that drug free film cross-linked by genipin （F0.1） possesses very low cytotoxicity to the tumor cells.The released VCR showed high in vitro inhibitive effects on HeLa,MCF-7 and A549 with different sensitivity on each line.Conclusions F0.1 film can achieve the release kinetics at a relatively constant speed.It has the potential for application as a carrier of anticancer drugs.