用碳二亚胺将明胶固定于壳聚糖膜表面,以改善壳聚糖的细胞相容性。采用MTT法评价膜材料对L929细胞的毒性。将壳聚糖膜(CS)、明胶固定化壳聚糖膜(CS—GEL)与人角膜上皮细胞(HCEC)体外复合培养,观察细胞的形态及生长情况,考察材料的细胞粘附率、材料对细胞活性与增殖的影响,并通过流式细胞术对细胞周期和凋亡进行分析。结果表明,CS-GEL具有良好的细胞粘附性,HCEC在CS—GEL上的生长和增殖情况优于CS,与CS相比,CS-GEL上HCEC的G1期比例下降,S期和G2期增加。明胶的引入缩短了细胞在膜表面的适应时间,使其尽快进入正常的细胞周期;同时表面固定的明胶可抑制细胞凋亡。因此,明胶固定化壳聚糖膜具有良好的体外细胞相容性,有望成为一种较好的角膜修复材料。
Gelatin was immobilized onto chitosan film via 1-ethyl-dimethylamino-propyl carbodiimide to improve the cytocompatibility. MTT assay was employed to evaluate the cytotoxicity with L929 cells. Human epithelial cells (HCEC) were co-cultured with chitosan film (CS) and gelatin-immobilized chitosan film (CS-GEL) in vitro. Cytocompatibility was investigated, such as cell morphology, adhesion, proliferation and the cell cycle and apoptosis. The results showed that HCEC were adhered on CS-GEL well, the growth and proliferation of HCEC on CS-GEL were obviously better than that of CS. Compared with CS, the cycle G1 of HCEC on CS-GEL reduced, while cycle S and G2 increased. The immobilization of gelatin onto chitosan films was in favor of the normal cell cycle due to the shortened cell adaptation period, and it could inhibit cell apoptosis as well. The gelatin-immobilized chitosan film had good biocompatibility with HCEC in vitro, it could be a potential biomaterial for corneal regeneration.