CAJ | 학술논문

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。
　　배경：관우골형태발생단백7작위자격인자유도세포성골적보도목전교소견。목적：관찰골막세포경골형태발생단백7유도후감성린산매적표체。 　　방법：취재우성인경골골막，상규세포배양법행골막세포체외배양，분위실험조화대조조，분별가입골형태발생단백7가성골세포배양보조제화단순성골세포배양보조제，상차현미경관찰골막세포형태특정급초미결구。매조분별재제7，14，21천설3개시간점，매개시간점설3개양본，채용감성린산매시제합법검측성골세포특이성표지물감성린산매표체정황。 　　결과여결론：골막세포경분조배양후，제7천시，실험조화대조조골막세포균유명현증식，감성린산매적가피검측출，단량불다，세포외형위사형，실험조비대조조검측적감성린산매수량초다；제14천시，실험조급대조조골막세포균현저증식，세포외형유사형변위관사형，실험조비대조조검측적감성린산매수량명현증다。제21천시，실험조급대조조골막세포균증식，기중실험조세포증식명현，세포외형위관사형，실험조비대조조검측적감성린산매수량현저증다。경과통계학분석유골형태발생단백7유도적골막세포적성골표지물감성린산매양성솔명현고우대조조(P<0.01)。제시골막세포구유량호적성골화재생능력,골형태발생단백7능유도골막세포가강감성린산매적표체，능유도골막세포향성골세포전화。
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 cel s after induced by bone morphogenetic protein-7 in vitro. METHODS:Periosteal cel s were obtained from adult tibial periosteum, and then the periosteal cel s were cultured by routine method in vitro. The cel s 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 cel s. 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 cel s in the experimental group and the control group was increased obviously, and the expression of alkaline phosphatase was detected but less. The cel s 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 cel s in the experimental group and the control group was increased obviously, the cel morphology was changed from spindle-shaped to wide spindle-shaped, and the expression of alkaline phosphatase in the experimental group was increased significantly when compared with the control group. After cultured for 21 days, the proliferation of periosteal cel s was detected in the experimental group and the control group, and the proliferation in the experimental group was more significant than that in the control group, the cel morphology was wide spindle-shaped, and the number of alkaline phosphatase in the experimental group was higher than that in the control group. Statistical analysis showed that the positive rate of osteogenic markers alkaline phosphatase of bone morphogenetic protein-7 induced periosteal cel s in the experimental group was higher than that in the control group (P<0.01). It suggested that periosteal cel s had the osteogenic and regeneration ability, the bone morphogenetic protein-7 could induce periosteal cel s, promote the expression of alkaline phosphatase, and could induce the periosteal cel s to transform into osteoblasts.