BMP-2体外诱导小鼠皮肤成纤维细胞向成骨细胞分化的可行性探讨  

Feasibility study on differentiation of skin fibroblasts into osteoblasts of mice induced by MMP-2 in vitro

作　　者：李波翰[1] 赵乾 刘斌[1] 高春正[1] 吴东进[1] 赵延英[1] 潘霄瀚 张颖哲 LI Bohan;ZHAO Qian;LIU Bin;GAO Chunzheng;WU Dongjin;ZHAO Yanying;PAN Xiaohan;ZHANG Yingzhe(The Second Hospital of Shandong University,Jinan 250033,China)

机构地区：[1]山东大学第二医院,济南250033 [2]山东大学附属济南市中心医院 [3]日本和歌山县立医科大学

出　　处：《山东医药》2018年第29期41-44,共4页Shandong Medical Journal

摘　　要：目的探讨骨形态发生蛋白-2(BMP-2)体外诱导小鼠皮肤成纤维细胞向成骨细胞分化的可行性。方法培养新生小鼠皮肤成纤维细胞,应用BMP-2诱导骨化,观察细胞形态变化,并应用茜素红染色和BMP免疫荧光进行细胞鉴定。结果应用含有BMP-2细胞培养液培养成纤维细胞,可见细胞形态学逐渐发生变化,具有成骨细胞形态学相关特征,茜素红染色阳性,具有成骨细胞生物学特性。茜素红染色半定量分析结果:含有BMP-2细胞培养液培养6 d后茜素红染色测定染色面积比例为0.3%;培养8 d后染色面积比例为0.75%;培养10 d后染色面积比例为2.42%;培养12 d后染色面积比例为17.61%。培养12 d后茜素红染色面积与其他三个时间点比较,差异有统计学意义(P均<0.05)。免疫荧光显示,含有BMP-2细胞培养液培养12 d后细胞能够有效表达BMP-2。结论BMP-2可诱导成纤维细胞在体外分化为成骨细胞。Objective To observe the feasibility of differentiation of skin fibroblasts into osteoblasts of mice induced by bone morphogenetic protein-2（MMP-2） in vitro. Methods The skin fibroblasts of newborn mice were cultured,and MMP-2 was used to induce ossification,and the cell morphological changes were observed. Alizarin red staining and BMP immunofluorescence were used for cell identification. Results After successful induction,the cellular morphology changed,and the morphological features of osteoblasts were observed,and alizarin red staining was positive. Semi-quantitative analysis results of Alizarin red staining： after 6 days of culture,the proportion of staining area was 0. 3% and 0. 45%,respectively,and the average staining area was 0. 375% ± 0. 075%; after 8 days of culture,the proportion of staining area was 0. 75% and 0. 75% respectively,and the average staining area was 0. 75%; after 10 days of culture,the proportion of staining area was 2. 42% and 2. 73%,respectively,and the average staining area was 2. 575% ± 0. 155%; after 12 days of culture,the proportion of staining area was 17. 61% and 15. 87%,respectively,and the average staining area was16. 74% ± 0. 87%. The difference between 12 days and the other three times was statistically significant（P 〈 0. 05）. Immunofluorescence showed that the cells,cultured in nutrient solution with BMP-2 after 12 days,were able to effectively express BMP-2. Conclusion Bone morphogenetic protein induces the differentiation of fibroblasts into osteoblasts in vitro.

关 键 词：成纤维细胞 成骨细胞 骨化 骨形态发生蛋白-2 

分 类 号：Q813[生物学—生物工程]