摘要
目的探讨骨形成蛋白-2（BMP-2）对OPG-/-小鼠骨髓间充质干细胞（BMsCs）体外定向诱导成骨的能力，评价BMP-2活性多肽的成骨诱导性及诱导成骨的剂量依赖性。方法取OPG-／-小鼠股骨，分离骨髓基质细胞进行体外培养，传至P，代时改用条件培养基，在15％FBS培养条件下培养基中分别加入：农度为0．3岭、0．2ug、0．1ug、0．05ug、0ug的BMP-2，并依次记为A、B、c、D和E组。细胞培养至1、3、5、7d，PNPP法测定碱性磷酸酶（ALP）、钙试剂盒测定细胞钙含量、酶联免疫法测定细胞上清中抗酒石酸酸性膦酸酶5b（TRACP5b）的表达，检测不同浓度BMP．2活性多肽体外诱导成骨的能力。结果ALP活性和钙含量检测显示，A组和B组较其他组增加明显（P〈0．05），但A、B组间比较差异无统计学意义（P〉0．05）。TRACP5b检测显示，A、B、c组第3、5和7日TRACP5b表达均低于D、E组（P〈0．05），但A和B组差异不显著（P〉0．05）。结论BMP-2活性多肽能有效地促进OPG-／-小鼠BMSCs向成骨方向分化，其诱导作用存在剂量依赖关系，最佳诱导剂量为0．2ug／ml。
ObjectiveTo investigate the effect of bone morphogenetic protein-2（BMP-2） on inducing OPG4- BMSCs differentiate into the osteogenesis in vitro, and to evaluate the effect of BMP-2 Bioactive Peptides inducing to osteoplast and dose dependent. Methods Got the femur of mice, separated the BMSCs and cultured in vitro. Followed by passaged the BMSCs to P1 using the conditioned medium, and then the BMSCs were added in the BMP-2 consistency of 0.3 ug, 0.2 ug, 0.1 ug, 0.05 ug, 0 ug separately cultured with 15% FBS culture medium, marked A, B, C, D, and E group in order. The 3rd, 5th, 7th day after cell culture, the expression of alkaline phosphatase was detected by PNPP method, and calcium content was detected by Calcium kit. The expression of tatrate-resistant acid phosphatase form 5 b in the cell supernatant detected by ELISA, To investigate the effect of different concentration bone morphogenetic protein-2 （BMP-2） on inducing OPG4- BMSCs differentiate into the osteogenesis in vitro. Results The activity of alkaline phosphatase and calcium content showed that group A and group B both increase higher than the other group （P〈0.05）. However no significant differences were detected be- tween the group A and B （P〉0.05）. TRACP5b displayed, TRACP5b expression in group A, B and C is lower than the other groups in the 3rd, 5th, 7th day （P〈0.05）, no significant differences were detected between the group A and B （P〉0.05）. Conclusions The BMP-2 had significant effect on inducing OPG- /- BMSCs differentiate into the osteogenesis in vitro. The effect dependent on the different concentration of bone morphogenetic protein-2（BMP-2）, the optimal concentration is 0.2 ug/ml.
出处
《实验动物与比较医学》
CAS
2012年第3期186-190,共5页
Laboratory Animal and Comparative Medicine
基金
[基金项目]新疆维吾尔自治区科技支疆项目资助（项目编号201091154）新疆医科大学第一附属医院组织工程专项基金资助（项目编号2009-ZZGC-04）
关键词
BMP-2
OPG-/-小鼠
骨髓间充质干细胞
诱导成骨
Bone morphogenetic protein-2（BMP-2）
OPG-/ mouse
Bone marrow derived stroma cell
Inducing into osteogenesis
