成纤维细胞生长因子1型受体-显性失活作用对骨诱导培养骨髓基质干细胞矿化的影响  

Effect of fibroblast growth factor receptors 1 dominant negative on mineralization of bone marrow stromal cells in bone induction culture

作　　者：李霞 徐文漭 简洪 李福兵 徐永清 

机构地区：[1]解放军昆明总医院骨科,全军创伤骨科研究所,650032 [2]解放军昆明总医院病理科,650032

出　　处：《中华创伤骨科杂志》2017年第9期801-805,共5页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金（81171734）;云南省创新团队基金（2009C1008）;国家临床重点专科建设项目

摘　　要：目的探讨成纤维细胞生长因子1型受体．显性失活作用（FGFRlDN）对骨诱导培养骨髓基质干细胞（BMSCs）成骨矿化的影响。方法取第3代BMSCs转染，根据细胞转染方法不同分为4组（n=24）：FGFRl-DN组[采用真核表达质粒pcDNA3．1（＋）-FGFRlDN转染]、FGFRl组【采用pcDNA3．1（＋）．FRFRl转染】、空载体转染组和未转染组。证实转染成功后，待细胞处于对数生长期时进行成骨诱导培养，连续培养21d，观察矿化结节形成情况并行茜素红染色，然后根据茜素红浓度标准曲线计算矿化物质的量。结果连续成骨诱导21d后，肉眼可见孔底圆形不透明钙化结节，经茜素红染色后呈红色，FGFRl-DN组BMSCs在对数生长期经成骨诱导培养，其成骨能力明显增加，而FGFRl组成骨能力减弱。茜素红浓度含量FGFRl-DN组最高（1．33±O．19），FGFRl组最低（1．00±1．17），空载体转染组（1．20±0．16）和未转染组（1．17±0．17）介于二者之间，组间比较差异有统计学意义（P〈0．05）。结论FGFRl-DN在BMSCs分化的早期可以促进细胞增殖，于对数生长期进行骨诱导培养后实现了其在成骨细胞中促进矿化的作用，这为局部基因治疗骨折骨缺损提供了理论依据。Objective To investigate the mineralization impact of fibroblast growth factor receptors 1 dominant negative （FGFR1DN） on osteogenic induction culture of bone marrow stromal cells （BMSCs） . Methods The 3rd generation BMSCs were divided into 4 equal groups （ n = 24） . FGFR1-DN group was transfected by pcDNA3.1 （ ＋ ） -FGFR1DN, FGFR1 group by pcDNA3.1 （ ＋ ） -FRFR1, blank load group by pcDNA3.1 （ ＋ ） -blank vehicle and non-transfection group by nothing. After successful transfection was con- firmed when the cells were in the logarithmic phase, osteogenic induction culture was conducted continuously for 21 days. Mineralized nodule formation was observed by alizarin red staining. The amount of mineralized material was calculated according to the standard curve of alizarin red concentration. Results Continuous osteogenic induction for 21 days showed on the bottom of the hole visible round opaque calcified nodules after alizarin red staining. The BMSCs in the FGFR1-DN group induced in the logarithmic growth phase by os- teoblasts exhibited significantly increased osteogenic capacity while those in the FGFR1 group displayed di-minished osteogenic capacity. The concentration of alizarin red was the highest in the FGFR1-DN group （1.33 ±0. 19）, the lowest in the FGFRI group （1.00 ± 1. 17）, and moderate in the blank load group （1.20 ±0. 16） and non-transfection group （1.17 ±0. 17）, showing significant between-group differences （ P 〈 0.05）. Conclusions FGFR1-DN can promote cell proliferation in the early differentiation of BMSCs and also mineralization of osteoblasts in bone induction culture after the logarithmic growth phase. This may provide a hint for local gene Ireatment of bone defects.

关 键 词：骨髓细胞 成纤维细胞生长因子受体1 显性失活 矿化 

分 类 号：R68[医药卫生—骨科学]