摘要
目的观察以纤维蛋白胶(fibrinsealant,FS)为载体复合骨形成蛋白(bonemorphogeneticprotein,BMP)的注射型骨修复材料对体外培养的兔骨髓基质细胞(marrowstromalcells,MSCs)增殖和分化的影响,为其临床应用提供实验依据。方法取3日龄新西兰兔骨髓进行培养传代,采用第3代细胞进行研究。实验分为3组,实验组以含1μg/mlrhBMP-2注射型FS培养细胞;FS对照组以单纯FS培养细胞;空白对照组不作任何处理。采用细胞培养、组织化学及电镜等方法对各组兔MSCs的增殖、贴壁率、碱性磷酸酶(alkalinephosphatase,ALP)活性及染色、型胶原表达、超微结构及细胞在材料中的生长情况进行观察。结果各组细胞促增殖作用由强到弱依次是FS对照组→实验组→空白对照组,各组间差异均有统计学意义(P<0.05);对细胞贴壁率的影响总体由强到弱依次是FS对照组→实验组→空白对照组,各组间差异均有统计学意义(P<0.05)。各组ALP活性由强到弱依次是实验组→FS对照组→空白对照组,各组间差异均有统计学意义(P<0.05);各组细胞的型胶原表达水平由高到低依次是实验组→FS对照组→空白对照组,差异均有统计学意义(P<0.05)。扫描电镜观察,材料表面粗糙,有微孔存在,FS对照组、实验组细胞与材料融合生长。透射电镜观察实验组细胞向成骨细胞分化程度高,但细胞增殖活性较FS对照组稍差;FS对照组细胞向成骨细胞分化的程度较实验组低,细胞增殖活性较好;空白对照组的细胞增殖活性较差,胞外基质少。结论以FS为载体复合BMP的注射型骨修复材料可显著提高MSCs向成骨细胞方向的分化水平,但对MSCs促增殖作用不明显。
Objective To investigate the effect of the injectable osteoinductive material with fibrin sealant（FS） as a carrier compounded with bone morphogenetic protein （BMP） on the proliferation and differentiation of marrow stromal cells （MSCs） towards osteoblasts and to provide the experimental foundation for the clinical application. Methods MSCs were extracted and cultured from bone marrow of the 3-day-old rabbit, and the third generation cultured MSCs were studied. The experiment included the experimental group （FS,including 1 μg/ml rhBMP-2）, FS control group（FS）and blank control group （no material）. The proliferation rate, the adhesive rate, the expression of the collagen Ⅰ and alkaline phosphatase, cell growth condition in the material and the ultrastructure of MSCs were investigated by electron microscopy, histochemistry and cell culture. Results The proliferation rate and the adhesive rate of MSCs in experimental group was significantly higher than those in blank control group ,but lower than those in FS control group （P〈0. 05）. The expression level of the collagen I and alkaline phosphatase in the experimental group was significantly higher than those in all control groups （P〈0. 05）. Scanning electron microscope showed that the surface of material was rough and had many pores and that cell and material mixed. Transmission electron microscope showed that MSCs of the experimental group were mostly of the phenotype of osteoblasts with relatively low proliferation activity and high differentiation degree toward osteoblasts and with plenty of extracellular matrix and collagen fibers. MSCs of FS control group had low differentiation degree toward osteoblasts with few extracellular matrix and collagen fibers and high proliferation activity. MSCs of blank control group had low differentiation degree toward osteoblasts with few extracellular matrix and collagen fibers, and low proliferation activity. Conclusion The injectable osteoinductive material with fibrin sealant as a carrier compounded with BMP could significantly accelerate the differentiation of MSCs towards osteoblasts. But it could not significantly accelerate the proliferation activity of MSCs.
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2007年第1期70-75,共6页
Chinese Journal of Reparative and Reconstructive Surgery
基金
全军医学科研计划十五课题资助项目(02M009)~~
关键词
骨髓基质细胞
纤维蛋白胶
骨形成蛋白
细胞培养
兔
Marrow stromal cells Fibrin sealant Bone morphogenetic protein Cell culture Rabbits
