摘要
目的:观察兔骨髓基质细胞经rhBMP-2与bFGF刺激后,其VEGF因子表达及成骨潜能的变化。方法:取兔双侧股骨骨髓基质细胞,采用骨髓基质细胞体外培养技术,单独或联合以rhBMP-2、bFGF刺激细胞。细胞培养5 d后,进行细胞形态(HE染色)、增殖情况(细胞记数法与MTT法)、碱性磷酸酶(改良钙钴法染色)、成骨结节(图像分析)、VEGF阳性细胞率(免疫组化染色)等项目的检测。结果:①联合应用rhBMP-2、bFGF在细胞记数、ALP活性、矿化面积百分率、VEGF阳性细胞率4个检测项目上优于单独应用rhBMP-2或bFGF,差别有统计学意义。②分开联合应用rhBMP-2、bFGF优于同时应用rhBMP-2或bFGF。结论:合理的联合使用rhBMP-2和bFGF,不仅可促进骨髓基质细胞的快速增殖及向成骨细胞转化,还可促进促血管内皮细胞增生的重要介质VEGF的表达。此项技术可应用于骨组织工程,促进工程化骨在体内的存活。
Objective ：To investigate the effects of rhBMP-2 （Recombinant Human Bone Morphogenetie Protein-2 ） and bFGF （Basic Fibroblast Growth Factor） on the VEGF（ Vascular Endothelial Growth Factor） expression and osteogenie potential of marrow stromal cells. Methodes：The marrow stromai cells of the femur of the rabbit were procured and cultured, The samples were treated with rhBMP-2 or （and） bFGF. After culturing for five days, the effects were observed by histopathologieal analysis, cells multiplication, alkaline phosphatase activities detection,the computer analysis of bone clusters, detection of ratio VEGF-positive eells（Immunoreaetivity）. Result：（1）The effects of combined use of BMP-2 and bFGF on marrow stromal cells were better than that of using rhBMP-2 or bFGF singly, there are statistical difference between them. （2）The effects of combined use of rhBMP-2 and bFGF on marrow stromal cells at the different time were better than that of combined use of rhBMP-2 and bFGF at the same time, there are statistical difference between them. Conclusions ： A rational combined use of rhBMP-2 and bFGF can not only enhances multiplication ability and osteogenie potentials of marrow stromal cells, but also promotes VEGF expression. The skill can be used in the field of tissue engineering of bone, which will be helpful for keeping the artificial bone alive in human＇s body.
出处
《中医正骨》
2009年第3期6-9,共4页
The Journal of Traditional Chinese Orthopedics and Traumatology
