可注射性纤维蛋白胶对兔骨髓基质细胞增殖和分化的影响

The effect of injectable fibrin sealant on the proliferation and differentiation of marrow stromal cell of rabbit

目的观察纤维蛋白胶(FS)对体外培养的兔骨髓基质细胞(MSCs)增殖和分化的影响。方法实验分两组:含有FS的实验组和不加任何材料的对照组。采用细胞培养、组织化学及电镜等方法对两组MSC的增殖活性、贴壁率、碱性磷酸酶(ALP)表达、Ⅰ型胶原表达、超微结构及细胞在材料中的生长情况进行研究。结果①实验组MSCs的增殖活性和贴壁率明显高于对照组(P<0.05)。②实验组MSCs的ALP活性显著高于对照组(P<0.05)。实验组第7天时的ALP活性显著高于第5天时本组的ALP活性(P<0.05)。实验组MSCs的Ⅰ型胶原表达水平显著高于对照组(P<0.05)。③扫描电镜观察:实验组MSCs与材料融合生长。透射电镜观察:实验组的MSCs细胞增殖活性好,向成骨细胞方向分化水平高,而对照组MSCs的细胞增殖活性相对较低,分化相对较差。结论FS生物相容性好,对MSCs增殖和向成骨细胞方向的分化均有明显促进作用,可作为骨组织工程材料的注射型载体进行进一步研究。

Objective To investigate the effect of the injectable fibrin sealant（FS） on the proliferation and differentiation of marrow stromal ceils（MSCs） of rabbit. Methods The specimens were divided into experiment group and control group （no any material）. The influence of experiment group and control group on the proliferation rate, the adhesive rates, the expression of the type I collagen and alkaline phosphatase（ALP） of 5th day and 7th day cultured MSCs, ceil growth condition in the material and the uhrastructure of the third generation rabbit MSCs were investigated individually with transmission and scanning electron microscopy, histochemistry, ALP and HE staining. Results （1） The proliferation rates and the adhesive rates of rabbit MSCs in experiment group was significantly higher than in control group（ P 〈 0. 05 ）. （2） The expression levels of the type I collagen and alkaline phosphatase in experiment group was significantly higher than the expression level in control groups（ P 〈 0. 05 ）. The 7th day expression level of alkaline phosphatase was significantly higher than the 5th day expression level in experiment group（ P 〈 0.05 ）. （3） Scanning electron microscope showed that ceils mixed and grew together with FS. Transmission electron microscope showed that MSCs of experiment group had high proliferation activity and high differentiation degree toward osteoblast, and that MSCs of control group had low differentiation degree toward osteoblast and low proliferation activity. Conclusions FS has very good biocompatibility and could significantly accelerate the proliferation and differentiation towards osteoblasts of marrow stromal ceil of rabbit. FS is a good alternative for the further study as the injectable cartier for bone tissue engineering material.