Di Palma Fabrice, Guignandon Alain, Chamson Annette, Lafage-Proust Marie-Hélène, Laroche Norbert, Peyroche Sylvie, Vico Laurence, Rattner Aline
Laboratoire de Biologie du Tissu Osseux, Faculté de Médecine 15 rue Ambroise Paré, Université Jean Monnet, Equipe INSERM E366, Saint-Etienne 42023, Cedex 02, France.
Biomaterials. 2005 Jul;26(20):4249-57. doi: 10.1016/j.biomaterials.2004.10.041.
In a previous study we demonstrated that MG-63 cells cultured on Ti-6Al-4V discs covered by alumina ceramic and submitted to intermittent mechanical strain (IMS) presented morphological alteration associated with enhanced differentiation. Here we examine how the mechanical response of osteoblasts can be modulated by the nature of the substrate. MG-63 cells were cultured on four materials: polystyrene and Ti-6Al-4V (average roughness = 0.48 microm) as smooth substrates; Ti-6Al-4V (average roughness = 5.76 microm) and Ti-6Al-4V covered with alumina (average roughness = 5.21 microm) as rough substrates. Mechanical strains were applied for 15 min, three times a day for 1-5 days with a 600 microstrains magnitude and a 0.25 Hz frequency. IMS stimulated alkaline phosphatase activity by 25-35% on all substrates and had no effect on cell growth on either substrate. Fibronectin (FN) was chosen as representative of cell-matrix interaction. FN production was increased by 60% after 1 day of stretching only on alumina-coated discs. FN organization examined on smooth substrates was affected by 5 days of IMS, showing a thickening of the fibres. The same modifications induced by IMS were previously observed on alumina-covered discs. Vinculin expression was not affected by IMS whatever the substrate. Cell-cell interactions were determined by N-cadherin immunoblotting. N-cadherin expression was increased by IMS specifically on rough substrates. Our results suggest that the nature of the surface did not influence the up-regulation of alkaline phosphatase activity induced by IMS, but modulates specifically cell-substrate as well as cell-cell interactions in response to IMS.
在之前的一项研究中,我们证明了在覆盖有氧化铝陶瓷的Ti-6Al-4V圆盘上培养并承受间歇性机械应变(IMS)的MG-63细胞呈现出与分化增强相关的形态学改变。在此,我们研究成骨细胞的机械反应如何受到底物性质的调节。MG-63细胞在四种材料上培养:聚苯乙烯和Ti-6Al-4V(平均粗糙度 = 0.48微米)作为光滑底物;Ti-6Al-4V(平均粗糙度 = 5.76微米)和覆盖有氧化铝的Ti-6Al-4V(平均粗糙度 = 5.21微米)作为粗糙底物。施加机械应变15分钟,每天三次,持续1 - 5天,应变幅度为600微应变,频率为0.25赫兹。IMS在所有底物上均使碱性磷酸酶活性提高了25 - 35%,且对任何一种底物上的细胞生长均无影响。选择纤连蛋白(FN)作为细胞 - 基质相互作用的代表。仅在氧化铝涂层圆盘上拉伸1天后,FN产量增加了60%。在光滑底物上检测到的FN组织受到5天IMS的影响,表现为纤维增粗。之前在氧化铝覆盖的圆盘上也观察到了IMS诱导的相同改变。无论底物如何,纽蛋白表达均不受IMS影响。通过N - 钙黏蛋白免疫印迹法测定细胞 - 细胞相互作用。IMS特异性地在粗糙底物上增加了N - 钙黏蛋白的表达。我们的结果表明,表面性质不影响IMS诱导的碱性磷酸酶活性上调,但会特异性地调节响应IMS的细胞 - 底物以及细胞 - 细胞相互作用。