纤维粘连蛋白与碱性成纤维细胞生长因子对成骨细胞黏附的协同刺激作用

SYNERGISTIC EFFECT OF FIBRONECTION AND BASIC FIBROBLAST GROWTH FACTOR ON OSTEOBLAST ADHESION EFFICIENCY

目的研究纤维粘连蛋白(fibronection,FN)与碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)单独或联合应用时对成骨细胞在生物衍生骨支架上黏附的影响,并探索其潜在机制。方法取第3代成骨细胞,以bFGF(0.1、1、10和100ng/ml)刺激,接种于FN(0.1、1、10和100μg/ml)或多聚赖氨酸(Polylysine)修饰的生物衍生骨支架上,MTT法检测组间细胞黏附效率差异,电镜观察细胞密度与形态。GRGDS肽封闭后再次重复上述步骤。结果单独应用FN与bFGF均可增加成骨细胞在生物衍生骨支架上的黏附效率(P<0.05),联合应用时,产生的效应显著增强,两者存在交互作用(P<0.01)。而Polylysine与bFGF无此交互作用(P>0.05)。电镜观察发现,两者联合应用时细胞密度、铺展效果均优于单独应用。应用GRGDS肽后,FN联合bFGF产生的黏附促进作用被显著阻断。结论bFGF与FN对成骨细胞在生物衍生骨三维支架上的黏附存在协同刺激作用,细胞黏附产生这一效应的机制很可能是由RGD-整合素α5β1途径来介导的,需深入研究探讨。

Objective To investigate effects of the basic fibroblast growth factor （bFGF） and fibronection （FN） on the osteoblast adhesion on the bio-derived bone. Methods The third generation of the osteoblast was treated with bFGF 0. 1, 1, 10, and 100 ng/ml, respectively, and then was seeded in the bio-derived bone, which had been modified with FN 0. 1, 1, 10, and 100 μg/ml, or Polylysine, respectively. The cell adhesion was measured by the MTT assay. The cell density and the cell appearance were observed by the scanning electron microscope. The abovementioned procedures were repeated by an application of the GRGDS peptide. Results Both FN and bFGF could enhance the osteoblast adhesion efficiency on the bio-derived bone （P 〈 0. 05）. However, the osteoblast adhesion efficiency could be significantly strengthened by a combined use of FN and bFGF. FN and bFGF had a significant synergistic effect in statistics （P〈0. 01）, but Polylysine and bFGF had no such synergistic effect （P〉 0. 05）. The combined use of FN and bFGF had a better effect on the cell density and the cell appearance than either of them when observed with the scanning electron microscope. Adhesion efficiency generated by the combined use of FN and bFGF was significantly blocked by the application of the GRGDS peptide. Conclusion The combined use of FN and bFGF has a significant synergistic effect on the osteoblast adhesion efficiency on the bio-derived bone. This effect is probably mediated by the RGD-integrin α5β1 pathway.