骨形态发生蛋白7诱导骨膜细胞碱性磷酸酶的表达

Bone morphogenetic protein-7 induces the expression of alkaline phosphatase in periosteal cells

背景:关于骨形态发生蛋白 7 作为刺激因子诱导细胞成骨的报道目前较少见。目的:观察骨膜细胞经骨形态发生蛋白 7 诱导后碱性磷酸酶的表达。方法:取材于成人胫骨骨膜,常规细胞培养法行骨膜细胞体外培养,分为实验组和对照组,分别加入骨形态发生蛋白 7 加成骨细胞培养辅助剂和单纯成骨细胞培养辅助剂,相差显微镜观察骨膜细胞形态特征及超微结构。每组分别在第 7,14,21 天设 3 个时间点,每个时间点设 3 个样本,采用碱性磷酸酶试剂盒法检测成骨细胞特异性标志物碱性磷酸酶表达情况。结果与结论:骨膜细胞经分组培养后,第 7 天时,实验组和对照组骨膜细胞均有明显增殖,碱性磷酸酶的可被检测出,但量不多,细胞外形为梭形,实验组比对照组检测的碱性磷酸酶数量稍多;第 14 天时,实验组及对照组骨膜细胞均显著增殖,细胞外形由梭形变为宽梭形,实验组比对照组检测的碱性磷酸酶数量明显增多。第 21 天时,实验组及对照组骨膜细胞均增殖,其中实验组细胞增殖明显,细胞外形为宽梭形,实验组比对照组检测的碱性磷酸酶数量显著增多。经过统计学分析由骨形态发生蛋白 7 诱导的骨膜细胞的成骨标志物碱性磷酸酶阳性率明显高于对照组(P < 0.01)。提示骨膜细胞具有良好的成骨和再生能力, 骨形态发生蛋白 7 能诱导骨膜细胞加强碱性磷酸酶的表达,能诱导骨膜细胞向成骨细胞转化。

BACKGROUND： The reports on bone morphogenetic protein-7 as a stimulating factor to induce osteogenic are relatively rare. OBJECTIVE： To study the expression of alkaline phosphatase of periosteal cells after induced by bone morphogenetic protein-7 in vitro. METHODS： Periosteal cells were obtained from adult tibial periosteum, and then the periosteal cells were cultured by routine method in vitro. The cells were divided into experimental group and control group, and then cultured with bone morphogenetic protein-7 plus osteoblast culture adjuvants and simple osteoblast culture adjuvants, respectively. The phase contrast microscope was used to observe the morphology and ultrastructure of periosteal cells. Each group was observed at 7, 14 and 21 days, and three samples were observed at each time point. Alkaline phosphatase kit was used to detect the expression of osteoblast-specific markers alkaline phosphatase. RESULTS AND CONCLUSION： After cultured for 7 days, the proliferation of periosteal cells in the experimental group and the control group was increased obviously, and the expression of alkaline phosphatase was detected but less. The cells were spindle in shape, while the expression of alkaline phosphatase in the experimental group was higher than that in the control group. After cultured for 14 days, the proliferation of periosteal cells in the experimental group and the control group was increased obviously, the cell morphology was changed from