摘要
[目的]通过体外细胞毒性试验,细胞与支架材料复合培养实验和体内植入试验,评价壳聚糖-藻酸盐支架材料的细胞相容性。[方法]通过冷冻干燥法制备具有纵行、平行排列通道的壳聚糖-藻酸盐支架材料。将大鼠骨髓间充质干细胞(bone marrow derived stromalcell,BMSCs)与其浸提液培养,采用MTT法检测培养1、3、5d时支架材料的细胞毒性。在体外将BMSCs与壳聚糖-藻酸盐支架材料复合培养3、5、7d后行扫描电镜检测,并将其植入急性脊髓半横断损伤模型中,6周后行免疫荧光检测BMSCs在材料中的生长与分化情况。[结果]MTT法检测示壳聚糖-藻酸盐支架材料的细胞毒性为0-1级。扫描电镜观察细胞在支架材料复合培养3d后即可粘附于支架材料表面,细胞呈一定方向排列。术后6周,Neurofilament200、NSE免疫荧光检测证实,支架材料内有大量BMSCs存活,部分向神经元样细胞分化。[结论]壳聚糖-藻酸盐支架具有良好的细胞相容性,有望成为一种较理想的神经组织工程支架材料。
[Objective]To evaluate the cytocompatibility of chitosan-alginate scaffolds through in vitro cytoxicity experiment,cell-material co-culture experiment and in vivo implantation test.[Method]Chitosan-alginate scaffolds with longitudinal,paralleled multi-channels were produced via freeze-drying. Bone marrow derived stromal cells （BMSCs） were cultivated with the leach liquor. The cytotoxicity of scaffold was analyzed using MTT assay after 1,3 and 5 days of culture.Scanning electron microscopy was conducted after 3,5 and 7 days of BMSCs culture on the chitosan-alginate scaffold in vitro. Immunofluorescence detection was performed after implanting BMSCs and chitosan-alginate scaffolds in vitro in acute hemi-transaction spinal cord injury.[Result]The cytotoxicity of chitosan-alginate scaffold was in grade 0-1. Scanning electron microscopic observation indicated that the cells adhered to and grew on the surface of scaffold,arranging in a directional manner after 3 days of co-culture. NF200 and NSE fluorescence detection proved that a great many BMSCs were survival in the scaffold after 6 weeks,and that some transplanted cells differentiated into neuron-like cells.[Conclusion]Chitosan-alginate scaffold has a satisfactory cytocompatibility and may be an ideal tissue engineering scaffold material.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2010年第10期836-839,共4页
Orthopedic Journal of China
基金
全军医学科学技术研究"十一五"计划课题项目(编号:06MA081)
关键词
组织工程
壳聚糖
藻酸盐
细胞相容性
tissue engineering
chitosan
alginate
cytocompatibility
