BMP-2与TNF-α对成骨细胞增殖和分化作用的比较分析

A comparative analysis of the effect of BMP-2 and TNF-alpha( TNF-α) on osteoblast proliferation and differentiation

目的通过BMP-2及TNF-α分别刺激原代培养的成骨细胞,探讨BMP-2及TNF-α对成骨细胞增殖和分化不同作用并进行比较分析。方法在无菌条件下手术切取Wistar(1周龄)大鼠颅骨并提取成骨细胞,原代细胞培养、传代、纯化、扩增并进行鉴定。采用BMP-2及TNF-α进行诱导,观察成骨细胞增殖,用利用MTT法检测其变化,利用PNPP偶氮法(4-硝基苯基磷酸二钠盐)检测成骨细胞碱性磷酸酶(ALP)活性。结果通过ALP染色和钙化的细胞实验均为阳性,证明体外培养扩增的细胞具有典型的成熟成骨细胞的特征。采用BMP-2及TNF-α来诱导大鼠成骨细胞与对照组的差异有统计学意义(P<0.05)。细胞增殖活性及ALP活性随BMP-2浓度的升高而增加,而随TNF-α浓度的升高而降低。结论 BMP-2转染后的成骨细胞增殖和分化功能都有所提高。而TNF-α对成骨细胞的增殖和分化均有抑制作用。对体外培养的成骨细胞,BMP-2和TNF-α分别起到促进和抑制作用。

Objective To investigate the effect of BMP-2 and TNF alpha（ TNF-α） on osteoblast proliferation and differentiation and a comparative analysis was to be done. Methods Wistar rat（ 1 week old） skull waschipped by the surgery done under sterile conditions and osteoblasts were extracted,cultured,passed,purified and amplified. Different concentrations of BMP-2 and TNF-α were used to induce rat osteoblasts. Methyl thiazolyl tetrazolium（ MTT） assay was used to detect the proliferation of osteoblasts,and PP-nitrophenyl phosphate（ PNPP） was used to detect the alkaline phosphatase（ ALP） activity of osteoblasts. Results Cells stained and calcified by ALP were both positive,which has proven that the cells cultured in vitro have the typical characteristics of mature osteoblasts. Rat osteoblast proliferation activity and ALP activity increased with the increase of BMP-2 concentration,but decreased with the increase of TNF-α concentration,and the differnce was statistically significant（ P〈0. 05）. Conclusions Transfection of BMP-2 can improve the proliferation and differentiation of osteoblasts. TNF-α has inhibitory effect on the proliferation and differentiation of osteoblasts. BMP-2 and TNF alpha respectively promote and inhibit in vitro culture of osteoblasts through different mechanisms.